Carbapenem derivatives

ABSTRACT

Compounds having potent antimicrobial effects over Gram-positive bacteria and Gram-negative bacteria, a high stability to β-lactamases and DHP-I and a high safety for the human body and a process for producing the same. Carbapenem compounds represented by the following general formula (I) or salts thereof: ##STR1## wherein the ring A represents a 3- to 7-membered non-aromatic ring containing at least one nitrogen atom and optionally having a substituent other than R 6  ; R 1  represents hydrogen or methyl; R 2  and R 5  are the same or different and each represents hydrogen or a protecting group of the hydroxyl group; R 3  represents hydrogen or a protecting group of the carboxyl group; R 4  represents hydrogen, lower alkyl or a protecting group of the amino group; R 6  represents: (1) hydrogen, (2) lower alkyl, optionally substituted by an optionally protected hydroxy, carbamoyl, formimidoyl, acetimidoyl or ##STR2## wherein R 7  and R 8  are the same or different and each represents hydrogen, lower alkyl, or a protecting group of the amino group), or (3) a protecting group of the amino group or a protecting group of the imino group; and m is 0 or 1. These compounds have antimicrobial effects and are useful as a drug.

This application is a 371 of PCT/US95/01299 filed Jun. 29, 1995.

FIELD OF THE INVENTION

This invention relates to novel carbapenem derivatives or salts thereofand a process for producing the same.

BACKGROUND OF THE INVENTION AND PRIOR ART

Since thienamycins were found in 1976 (JP-A 51-73191(1976), there havebeen made a number of researches and developments on carbapenemantibiotics.

Even today, studies are under way to synthesize a number of carbapenemcompounds which have potent and broad antimicrobial spectra overGram-negative bacteria and Gram-positive bacteria and are highly stableto many β-lactamases.

For example, Taura et al. (JP-A 6-157544(1994)) have reported carbapenemcompounds having a β-configuration methyl group, a(6,7-dihydro-5H-pyrazolo[1,2-α] [1,2,4] triazolium-6-yl)] thio group anda 1-substituted-hydroxyethyl group respectively at the 1-, 2- and6-positions of the carbapenem skeleton. In addition, a number ofcompounds have been reported by JP-A 6-1791(1994), JP-A 6-16671(1994),JP-A 5-339269(1993), etc.

JP-A 6-87858(1994) has disclosed carbapenem derivatives having anaminoalkylpyrrolidinylthio group at the 2-position of the carbapenemskeleton and being similar to the compounds of the present invention instructure. This patent generally indicates carbapenem compounds having a2-substituted-pyrrolidin-4-ylthio group as the side chain at the2-position of the carbapenem skeleton wherein the substituent is abranched alkylene group having a primary, secondary or tertiary aminogroup or an ammonio group at the terminus. However, it neither states ingeneral nor suggests any compound which has a cyclic amine- orlactam-substituted hydroxymethyl group as the side chain of thepyrrolidinylthio group at the 2-position of the carbapenem skeleton.

Frequent use of penicillin and cephalosporin antibiotics having broadantimicrobial spectra has caused a social problem of the appearance ofresistant bacteria, in particular, methicillin-resistant Staphylococcusaureus (MRSA) and resistant Pseudomonas aeruginosa. There has been knownno drug efficacious against these resistant bacteria and thus it isurgently required to develop novel chemicals therefor.

As described above, carbapenem antibiotics have potent and broadantimicrobial spectra over Gram-negative and Gram-positive bacteria andare stable to β-lactamases. Although these carbapenem antibiotics areefficacious against bacteria resistant to a number of penicillin andcephalosporin antibiotics, their antimicrobial activities on theseresistant bacteria are not always sufficient. The carbapenem antibioticsare poor in chemical stability and, furthermore, very easily metabolizedby dehydropeptidase-I (DHP-I) localized in the kidney, etc., and thuslose their antimicrobial activities within a short period of time invivo. These properties make the compounds not necessarily useful as adrug.

In addition, there remain some problems regarding safety for the humanbody (for example, side effects on the central nervous system andnephrotoxicity due to decomposition products) and an increase inresistant bacteria caused by the frequent use of carbapenem antibioticssuch as thienamycin and imipenem.

Although several carbapenem antibiotics are now employed in thetreatment of infectious diseases, there has not been developed acarbapenem antibiotic which has an excellent antimicrobial effect and abroad antimicrobial spectrum and is satisfactory in stability in vivo,safety for the human body (i.e., avoiding any toxicity), etc.

DISCLOSURE OF THE INVENTION

The present inventors have conducted extensive studies on the synthesisof carbapenem derivatives in order to solve the above-mentioned problemsand consequently found out that compounds represented by the followinggeneral formula (I), which have 2-substituted pyrrolidinylthio groupshaving various substituents at the 2-position, are useful as drugsbecause of their potent antimicrobial activities and high safety for thehuman body, thus completing the present invention.

Accordingly, the present invention relates to novel carbapenemderivatives represented by the following general formula (I) and beinguseful as antimicrobial agents, or salts thereof, and a process forproducing the same: ##STR3## wherein the ring A represents a 3- to7-membered ring containing at least one nitrogen atom and optionallyhaving a substituent other than R⁶ ; R¹ represents hydrogen or methyl;R² and R⁵ are the same or different and each represents hydrogen or aprotecting group of the hydroxyl group; R³ represents hydrogen or aprotecting group of the carboxyl group; R⁴ represents hydrogen, loweralkyl or a protecting group of the amino group; R⁶ represents: (1)hydrogen, (2) lower alkyl optionally substituted by optionally protectedhydroxy, carbamoyl, formimidoyl, acetimidoyl or ##STR4## (wherein R⁷ andR⁸ are the same or different and each represents hydrogen, lower alkyl,or a protecting group of the amino group), or (3) a protecting group ofthe amino group or a protecting group of the imino group; and m is 0 or1.

The present invention further provides an antimicrobial agent comprisingas the active ingredient the above-mentioned compound or apharmacologically acceptable salt thereof; the use of theabove-mentioned compound or salt thereof for producing an antimicrobialagent; a method for preventing or treating microbisms which comprisesadministering a pharmacologically efficacious dose of theabove-mentioned compound, or salt thereof, to a patient; and a medicinalcomposition which comprises a pharmacologically efficacious amount ofthe above-mentioned compound, or salt thereof, and pharmacologicallyacceptable carrier(s).

Now, the contents of the present invention and the terms, etc., usedherein will be described in detail.

First, the compounds of the present invention include optical isomersand stereoisomers on the basis of asymmetric carbon atoms at the 1-, 5-,6- and 8-positions of the carbapenem skeleton, as shown in the followingformula: ##STR5## wherein R¹ represents hydrogen or methyl; and *represents an asymmetric carbon atom. Although these isomers are allrepresented by a single formula for the sake of convenience, the presentinvention is not restricted to the formula given for the sake ofconvenience but involves all of the isomers and isomer mixtures on thebasis of each of these asymmetric carbons.

Further, there are isomers on the basis of the asymmetric carbon atom inthe substituent on the side chain at the 2-position of the carbapenemskeleton. The present invention involves all of these optical isomersand stereoisomers too.

From the viewpoint of antimicrobial effect, however, the compoundsrepresented by the following formula, or salts thereof, are preferable:##STR6## wherein the ring A represents a 3- to 7-membered ringcontaining at least one nitrogen atom and optionally having asubstituent other than R⁶ ; R¹ represents hydrogen or methyl; R² and R⁵are the same or different and each represents hydrogen or a protectinggroup of the hydroxyl group; R³ represents hydrogen or a protectinggroup of the carboxyl group; R⁴ represents hydrogen, lower alkyl or aprotecting group of the amino group; R⁶ represents: (1) hydrogen, (2)lower alkyl optionally substituted by optionally protected hydroxy,carbamoyl, formimidoyl, acetimidoyl or ##STR7## (wherein R⁷ and R⁸ arethe same or different and each represents hydrogen, lower alkyl, or aprotecting group of the amino group), or (3) a protecting group of theamino group or a protecting group of the imino group; and m is 0 or 1.

R¹ represents a hydrogen atom or a methyl group and methyl is preferablyused therefor.

R² and R⁵ are the same or different and each represents a hydrogen atomor a protecting group of the hydroxyl group. The protecting group of thehydroxyl group may be an arbitrary one without restriction, so long asit is a group commonly known as a protecting group of a hydroxyl groupin organic syntheses. Particular examples thereof include loweralkylsilyl groups such as trimethylsilyl and t-butyldimethyl-silylgroups; lower alkoxymethyl groups such as methoxymethyl and2-methoxyethoxymethyl groups; a tetrahydropyranyl group and the like;aralkyl groups such as benzyl, p-methoxybenzyl, 2,4-dimethoxybenzyl,o-nitrobenzyl, p-nitrobenzyl and trityl groups; acyl groups such asformyl and acetyl groups; lower alkoxycarbonyl groups such ast-butoxycarbonyl, 2-iodoethoxy-carbonyl and2,2,2-trichloroethoxycarbonyl groups; alkenyloxy-carbonyl groups such as2-propenyloxycarbonyl, 2-chloro-2-propenyloxycarbonyl,3-methoxycarbonyl-2-propenyloxycarbonyl, 2-methyl-2-propenyloxycarbonyl,2-butenyloxycarbonyl and cinnamyloxycarbonyl groups; andaralkyloxycarbonyl groups such as benzyloxycarbonyl,p-methoxybenzyloxycarbonyl, o-nitro-benzyloxycarbonyl andp-nitrobenzyloxycarbonyl groups.

R³ represents a hydrogen atom or a protecting group of the carboxylgroup.

The protecting group of the carboxyl group may be an arbitrary onewithout restriction, so long as it is a group commonly known as aprotecting group of a carboxyl group in organic syntheses. Particularexamples thereof include linear and branched lower alkyl groups having 1to 4 carbon atoms such as methyl, ethyl, isopropyl and t-butyl groups;halogenated lower alkyl groups such as 2-iodoethyl and2,2,2-trichloro-ethyl groups; lower alkoxymethyl groups such asmethoxymethyl, ethoxymethyl and isobutoxymethyl groups; lower aliphaticacyloxymethyl groups such as butyryloxymethyl and pivaloyloxy-methylgroups; 1-(lower alkoxy)carbonyloxyethyl groups such as1-methoxycarbonyloxyethyl and 1-ethoxycarbonyloxy-ethyl groups; aralkylgroups such as benzyl, p-methoxybenzyl, o-nitrobenzyl and p-nitrobenzylgroups; a benzhydryl group and a phthalidyl group.

R⁴ represents a hydrogen atom, a lower alkyl group or a protecting groupof the amino group.

The lower alkyl group means a linear or branched alkyl group having 1 to6 carbon atoms such as methyl, ethyl, n-propyl, i-propyl, n-butyl,i-butyl, sec-butyl, t-butyl, n-pentyl, i-pentyl, sec-pentyl, t-pentyl,neopentyl, 1-methylbutyl, 2-methylbutyl, 1,1-dimethylpropyl,1,2-dimethylpropyl, n-hexyl, i-hexyl, 1-methylpentyl, 2-methylpentyl,3-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl,1,3-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl,2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl,1-ethyl-1-methylpropyl and 1-ethyl-2-methylpropyl groups.

The protecting group of the amino group may be an arbitrary one withoutrestriction, so long as it is a group commonly known as a protectinggroup of an amino group in organic syntheses. Particular examplesthereof include optionally substituted lower alkanoyl groups such asformyl, acetyl, chloroacetyl, dichloroacetyl, propionyl, phenylacetyl,phenoxyacetyl and thienylacetyl groups; optionally substituted loweralkoxycarbonyl groups such as benzyloxycarbonyl, t-butoxycarbonyl andp-nitrobenzyloxycarbonyl groups; substituted lower alkyl groups such asmethyl, t-butyl, 2,2,2-trichloroethyl, trityl, p-methoxybenzyl,p-nitrobenzyl, diphenylmethyl and pivaloyloxymethyl groups; substitutedsilyl groups such as trimethylsilyl and t-butyldimethylsilyl groups; andoptionally substituted benzylidene groups such as benzylidene,salicylidene, p-nitrobenzylidene, m-chlorobenzylidene,3,5-di(t-butyl)-4-hydroxybenzylidene and 3,5-di(t-butyl)benzylidenegroups.

Such a protecting group can be eliminated by any conventional methodsuch as hydrolysis or reduction depending on the type thereof.

R⁶ represents: (1) a hydrogen atom, (2) a lower alkyl group optionallysubstituted by optionally protected hydroxyl, carbamoyl, formimidoyl,acetimidoyl or ##STR8## (wherein R⁷ and R⁸ are the same or different andeach represents a hydrogen atom, a lower alkyl group or a protectinggroup of the amino group), or (3) a protecting group of the amino groupor a protecting group of the imino group.

As the examples of the protecting group of the hydroxyl group in theoptionally protected hydroxyl group, citation can be made of theprotecting groups of a hydroxyl group described above.

The lower alkyl group and the protecting group of the amino group in R⁷and R⁸ are each the same as those defined above.

The protecting group of the amino group or the protecting group of theimino group in R⁶ may be an arbitrary one without restriction, so longas it is a group commonly known as a protecting group of an amino groupor a protecting group of an imino group in organic syntheses. Particularexamples thereof include optionally substituted lower alkanoyl groupssuch as formyl, acetyl, chloroacetyl, dichloroacetyl, propionyl,phenylacetyl, phenoxyacetyl and thienylacetyl groups; optionallysubstituted lower alkoxycarbonyl groups such as benzyloxycarbonyl,t-butoxycarbonyl and p-nitrobenzyloxy-carbonyl groups; substituted loweralkyl groups such as methyl, t-butyl, 2,2,2-trichloroethyl, trityl,p-methoxy-benzyl, p-nitrobenzyl, diphenylmethyl and pivaloyloxymethylgroups; substituted silyl groups such as trimethyl-silyl andt-butyldimethylsilyl groups; and optionally substituted benzylidinegroups such as benzylidene, salicylidene, p-nitrobenzylidene,m-chlorobenzylidene, 3,5-di(t-butyl)-4-hydroxybenzylidene and3,5-di(t-butyl)benzylidene groups.

Such a protecting group can be eliminated by any conventional methodsuch as hydrolysis or reduction depending on the type of the protectinggroup.

Accordingly, particular examples of R⁶ include, in addition to thehydrogen atom, lower alkyl groups substituted by optionally protectedhydroxyl groups such as hydroxymethyl, hydroxyethyl, hydroxypropyl andhydroxybutyl groups; lower alkyl groups substituted by carbamoyl groupssuch as carmaboylmethyl, carbamoylethyl, carbamoylpropyl andcarbamoylbutyl groups; lower alkyl groups substituted by formimidoylgroups such as formimidoylmethyl, formimidoylethyl, formimidoylpropyland formimidoylbutyl groups; lower alkyl groups substituted byacetimidoyl groups such as acetimdoylmethyl, acetimidoylethyl,acetimidoylpropyl and acetimidoylbutyl groups; substituted amino-loweralkyl groups such as aminomethyl, aminoethyl, aminopropyl, aminobutyl,N-methylaminomethyl, N-methylaminoethyl, N-methylaminopropyl,N-methylaminobutyl, N,N-dimethylaminomethyl, N,N-dimethylamino-ethyl,N,N-dimethylaminopropyl, N,N-dimethylaminobutyl,N,N-ethylmethylaminomethyl, N,N-ethylmethylaminoethyl,N,N-ethylmethylaminopropyl and N,N-ethylmethylaminobutyl groups; and theabove-mentioned protecting groups of an amino group and the protectinggroups of an imino group, though the present invention is not restrictedthereto.

m is 0 or 1.

The type of the salt is not restricted and examples thereof includeinorganic acid addition salts such as hydrochloride, sulfate, carbonate,bicarbonate, hydrobromide and hydriodide; organic carboxylic acidaddition salts such as acetate, maleate, lactate, tartrate andtrifluoroacetate; organic sulfonic acid addition salts such asmethanesulfonate, hydroxymethanesulfonate, hydroxyethanesulfonate,benzenesulfonate, toluenesulfonate and taurine salt; amine additionsalts such as trimethylamine salt, triethylamine salt, pyridine salt,procaine salt, picoline salt, dicyclohexylamine salt,N,N'-dibenzylethylenediamine salt, N-methyl-glucamine salt,diethanolamine salt, triethanolamine salt,tris(hydroxy-methylamino)methane salt and phenethylbenzylamine salt;alkali metal addition salts such as sodium salt and potassium salt;alkaline earth metal addition salts such as magnesium salt and calciumsalt; and amino acid addition salts such as arginine salt, lysine salt,serine salt, glycine salt, aspartate and glutamate.

The term "pharmacologically acceptable salt" as used herein refers tothe conventional ones commonly employed in the production of drugs.

The ester in the above general formula (I) refers to an ester at thecarboxyl group at the 3-position of the carbapenem skeleton containingan ester group which is physiologically acceptable and hydrolyzableunder physiological conditions. Examples thereof include C₁₋₆ alkyl,benzyl, 4-methoxybenzyl, alkanoyloxyalkyl (for example, acetoxymethyl,propionyloxymethyl and pivaloxymethyl), alkoxy-carbonyloxyalkyl (forexample, methoxycarbonyloxymethyl, ethoxycarbonyloxymethyl and2-methoxycarbonyloxyethyl) and 5-(methyl-2-oxy-1,3-dioxo-4-yl)-methylgroups.

The ring A represents a 3- to 7-membered ring containing at least onenitrogen atom and optionally having a substituent other than R⁶.

Examples of the 3- to 7-membered ring containing at least one nitrogenatom include aziridine, azetidine, pyrrolidine, piperidine,homopiperidine, imidazolidine, pyrazolidine, piperazine, pyrroline,imidazoline and pyrazoline rings.

The expression "optionally having a substituent other than R⁶ " meansoptionally having one or more substituents, for example, a hydrogenatom, a hydroxyl group, a thiol group, halogen atoms, and nitrile,azido, alkyl, alkenyl, alkynyl, alkoxy, halogenoalkyl, guanidino,formimidoyl, acetimidoyl, carbamoyl, thiocarbamoyl, carbamoylalkyl,carbamido, alkanoyl, amino, alkylamino, dialkylamino, aminoalkylcarboxy, alkoxycarbonyl, alkoxycarbonylalkyl, aminoalkyl-aminoalkyl,alkylcarbonyloxy, ammonium, cycloalkyl, cycloalkenyl, phenyl, alkylthio,phenylthio, benzyl, benzoyl and halogenoaryl groups.

The present inventors have found out that carbapenem derivatives having,as the side chain of the pyrrolidinylthio group at the 2-position of thecarbapenem skeleton, a group represented by the following generalformula, have excellent antimicrobial activities, thus completing thepresent invention: ##STR9## wherein the ring A represents a 3- to7-membered ring containing at least one nitrogen atom and optionallyhaving a substituent other than R⁶ ; R⁵ represents hydrogen or aprotecting group of the hydroxyl group; R⁶ represents: (1) hydrogen, (2)lower alkyl optionally substituted by optionally protected hydroxy,carbamoyl, formimidoyl, acetimidoyl or ##STR10## (wherein R⁷ and R⁸ arethe same or different and each represents hydrogen, lower alkyl, or aprotecting group of the amino group), or (3) a protecting group of theamino group or a protecting group of the imino group; and m is 0 or 1.

The compounds of the present invention are characterized by having thestructures where the pyrrolidinylthio group, which is the side chain atthe 2-position of the carbapenem skeleton, has a cyclic amine- orlactam-substituted hydroxymethyl group at the 2-position of thepyrrolidine ring. Examples of the cyclic amine- or lactam-substitutedhydroxymethyl group include aziridinylhydroxymethyl,azetidinylhydroxymethyl, pyrrolidinylhydroxymethyl,piperidinylhydroxymethyl, homopiperidinylhydroxymethl,2-aziridinonylhydroxymethyl, 2-azetidinonylhydroxymethyl,2-pyrrolidinonylhydroxymethyl, 2-piperidinonylhydroxymethyl and2-homopiperidinonyl-hydroxy-methyl groups, each optionally havingsubstituent(s) other than R⁶.

Accordingly, preferable examples of the carbapenem derivatives of thepresent invention include the compound represented by the followingformula, and salts and esters thereof: ##STR11## the compoundrepresented by the following formula, and salts and esters thereof:##STR12## the compound represented by the following formula, and saltsand esters thereof: and ##STR13## the compound represented by thefollowing formula, and salts and esters thereof: ##STR14##

Next, a process for producing the compounds of the present inventionwill be described.

To produce a compound represented by the following general formula, or asalt thereof: ##STR15## (wherein the ring A represents a 3- to7-membered ring containing at least one nitrogen atom and optionallyhaving a substituent other than R⁶ ; R¹ represents hydrogen or methyl;R² and R⁵ are the same or different and each represents hydrogen or aprotecting group of the hydroxyl group; R³ represents hydrogen or aprotecting group of the carboxyl group; R⁴ represents hydrogen, loweralkyl or a protecting group of the amino group; R⁶ represents: (1)hydrogen, (2) lower alkyl optionally substituted by optionally protectedhydroxy, carbamoyl, formimidoyl, acetimidoyl or ##STR16## (wherein R⁷and R⁸ are the same or different and each represents hydrogen, loweralkyl, or a protecting group of the amino group), or (3) a protectinggroup of the amino group or a protecting group of the imino group; and mis 0 or 1), a compound represented by the following general formula(III), or a reactive derivative thereof: ##STR17## (wherein R¹represents hydrogen or methyl; R² represents hydrogen or a protectinggroup of the hydroxyl group; and R³ represents hydrogen or a protectinggroup of the carboxyl group) is reacted in the presence of a base with amercaptan represented by the following general formula: ##STR18##(wherein the ring A represents a 3- to 7-membered ring containing atleast one nitrogen atom and optionally having a substituent other thanR⁶ ; R⁴ represents hydrogen, lower alkyl or a protecting group of theamino group; R⁵ represents hydrogen or a protecting group of thehydroxyl group; R⁶ represents: (1) hydrogen, (2) lower alkyl optionallysubstituted by optionally protected hydroxy, carbamoyl, formimidoyl,acetimidoyl or ##STR19## (wherein R⁷ and R⁸ are the same or differentand each represents hydrogen, lower alkyl, or a protecting group of theamino group), or (3) a protecting group of the amino group or aprotecting group of the imino group; and m is 0 or 1), optionallyfollowed by the deblocking reaction. Thus, the compounds represented bythe above general formula (I), or salts or esters thereof, can beproduced. After eliminating the protecting group of R³, R³ represents ahydrogen atom.

To synthesize compounds represented by the above general formula (II)having particularly excellent antimicrobial effects, intermediatesrepresented by the following general formulae are used as thesubstitutes respectively for the above-mentioned intermediates (III) and(IV): ##STR20##

More particularly, reactive derivatives of the compounds represented bythe general formula (III): ##STR21## (wherein R¹ represents hydrogen ormethyl; R² represents hydrogen or a protecting group of the hydroxylgroup; and R³ represents hydrogen or a protecting group of the carboxylgroup) can be obtained by the following method. Thus, the above compoundis reacted with an activating reagent in an inert organic solvent in thepresence of a base to thereby give a reactive derivative represented bythe following general formula: ##STR22## wherein R¹, R² and R³ are eachas defined above; and L represents a leaving group.

An arbitrary solvent may be used therefor, so long as it does notinhibit the reaction. Examples of such a solvent include diethyl ether,tetrahydrofuran, dioxane, dichloro-methane, 1,2-dichloroethane,chloroform, carbon tetrachloride, trichloroethylene, hexane, benzene,chlorobenzene, toluene, ethyl acetate, butyl acetate,1,2-dimethoxyethane, N,N-dimethylformamide, N,N-dimethylacetamide,dimethyl sulfoxide, hexamethyl-phosphoramide, acetone, acetonitrile,water and mixtures thereof.

As the base, use can be made of, for example, sodium carbonate,potassium carbonate, sodium hydride, potassium hydride,t-butoxypotassium, trimethylamine, triethylamine,N,N-diisopropylethylamine, N-methyl-morpholine, N-methyl-pyrrolidine,N-methylpiperidine, N,N-dimethylaniline,1,8-diazabicyclo[5,4,0]undec-7-ene (DBU), pyridine,4-dimethyl-aminopyridine, picoline, lutidine, quinoline andisoquinoline.

Examples of the activating reagent to be used in the reaction includeacid anhydrides such as trifluoroacetic anhydride, methanesulfonicanhydride, trifluoromethane-sulfonic anhydride and p-toluene-sulfonicanhydride; and acid chlorides such as methanesulfonyl chloride,p-toluenesulfonyl chloride and diphenyl chlorophosphate.

In the above general formula, L represents a leaving group. Examplesthereof include trifluoroacetoxy, methanesulfonyloxy,trifluoromethane-sulfonyloxy, p-toluenesulfonyloxy anddiphenoxyphosphoryloxy groups.

The reaction is effected generally within a temperature range of -40 to50° C., preferably -20 to 20° C.

The reaction between this reactive derivative and the mercaptan compoundrepresented by the general formula ##STR23## (wherein the ring Arepresents a 3- to 7-membered ring containing at least one nitrogen atomand optionally having a substituent other than R⁶ ; R⁴ representshydrogen, lower alkyl or a protecting group of the amino group; R⁵represents hydrogen or a protecting group of the hydroxyl group; R⁶represents: (1) hydrogen, (2) lower alkyl optionally substituted byoptionally protected hydroxy, carbamoyl, formimidoyl, acetimidoyl or##STR24## (wherein R⁷ and R⁸ are the same or different and eachrepresents hydrogen, lower alkyl, or a protecting group of the aminogroup), or (3) a protecting group of the amino group or a protectinggroup of the imino group; and m is 0 or 1) is effected in theabove-mentioned inert organic solvent with the use of a base.Subsequently, the obtained product is subjected to the deblockingreaction, if necessary. Thus the compound represented by the abovegeneral formula (I) or a salt or ester thereof can be obtained. Similarto the above-mentioned reaction, this reaction can be carried outgenerally within a temperature range of -40 to 50° C., preferably -20 to20° C. It is possible to deblock the obtained compound as such withoutbeing purified. However, it is preferable to purify the product by anyconventional method (for example, column chromatography with the use ofsilica gel, etc.) and then subject it to the deblocking reaction.

The protecting group may be eliminated by effecting, for example,reduction such as catalytic reduction or solvolysis under usualconditions, though the appropriate elimination method varies dependingon the type of the protecting group.

The dose of the antimicrobial agent of the present invention variesdepending on the severity of the conditions, age, sex and weight of thesubject, the administration route, the disease, etc. Usually, it isadministered to an adult in a dose of from 1 to 1,000 mg per day in oneto several portions.

The administration route of the antimicrobial agent of the presentinvention is not particularly restricted. Namely, it can be orally orparenterally administered by any method commonly employed in the art.

The compounds of the present invention can be processed intopharmaceutical preparations by the conventional methods with the use offillers, binders, lubricants, colorants, corrigents, etc. commonlyemployed in the art together with, if necessary, stabilizers,emulsifiers, sorbefacients, surfactants, etc.

To process the compounds of the present invention into pharmaceuticalpreparations, use may be made of, in particular, DHP-I inhibitors suchas cilastatin.

The compounds of the present invention exhibit excellent antimicrobialactivities on various Gram-positive and Gram-negative bacteria. Toillustrate the usefulness of the compound of the present invention, theantimicrobial activities thereof on, in particular, P. aeruginosa, whichhas brought about clinical problems in recent years, will be givenhereinbelow.

                  TABLE 1                                                         ______________________________________                                         ##STR25##                                                                    MIC (μg/ml)                                                                ______________________________________                                        S. aureus E31295     0.1                                                      E. coli CS109        0.05                                                     H. influenzae IID1638                                                                              0.2                                                      P. aeruginosa E03763 0.8                                                      P. aeruginosa NCTC10490                                                                            0.1                                                      P. aeruginosa PA01   0.2                                                      P. aeruginosa E03441/SKR2-14                                                                       0.1                                                      P. aeruginosa E03441/SKR2/I-2                                                                      0.4                                                      P. aeruginosa E03441/WT                                                                            0.2                                                      P. aeruginosa E03441/SKR2                                                                          0.2                                                      P. aeruginosa E03441/R24                                                                           3.13                                                     P. aeruginosa E03402 0.1                                                      ______________________________________                                    

As the above Table 1 shows, the compound of the present invention has anexcellent antimicrobial effect and, therefore, is useful in theprevention and treatment of various microorganisms.

To further illustrate the present invention in greater detail, and notby way of limitation, the following Examples will be given. In theseExamples, ¹ H NMR spectra were measured by using a Varian FT NMR (400MHz) spectrometer.

EXAMPLES Production Example 1

(2S,4R)-N-tert-Butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butyldimethylsilyl-2-oxoazetidin-3-ylhydroxymethyl)-pyrrolidine##STR26##

At -30° C., a 1.6 M solution of n-butyllithium in hexane (10.6 ml, 17.0mmol) was added to a solution of diisopropyl-amine (2.6 ml, 19 mmol) intetrahydrofuran (50 ml). After stirring at the same temperature for 20minutes, the mixture was cooled to -78° C. Into the solution was droppeda solution of N-tert-butyldimethylsilyl-2-oxoazetidine (2.62 g, 14.2mmol) in tetrahydrofuran (15 ml) and the resulting mixture was stirredat -78° C. for 20 minutes. Into the solution was dropped a solution of(2S,4R)-N-tert-butoxy-carbonyl-4-tert-butyldimethylsiloxy-2-formyl-pyrrolidine(4.66 g, 14.2 mmol) in tetrahydrofuran (10 ml) and the resulting mixturewas stirred at the same temperature for 40 minutes. To the reactionmixture was added a saturated aqueous solution of ammonium chloride (15ml) and the mixture was returned to room temperature. Then water (50 ml)was added to the reaction mixture followed by extraction with diethylether (150 ml). The organic layer was washed successively with 1 Nhydrochloric acid, water, a saturated aqueous solution of sodiumhydrogencarbonate and a saturated aqueous solution of sodium chloride.Next, the organic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography [Wakogel C-200, 100 g, hexane-ethyl acetate(9:1 to 4:1)] to thereby give the target compound (3.38 g, 35.5%).

NMR(CDCl₃) δ: 0.05, 0.06(6H, each s), 0.20, 0.23(6H, each s), 0.86(9H,s), 0.95, 0.96(9H, each s), 1.46(9H, s), 1.70˜2.26(2H, m), 3.15˜3.60(5H,m), 3.76˜4.46(3H, m), 5.11(1H, d, J=9 Hz)

Production Example 2

(2S,4R)-N-tert-Butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-oxoazetidin-3-ylhydroxymethyl)pyrrolidine##STR27##

Under ice-cooling, acetic acid (0.52 ml, 9.1 mmol) and 1 M solution oftetrabutylammonium fluoride in tetrahydrofuran (9.1 ml, 9.1 mmol) wereadded to a solution of(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butyl-dimethylsilyl-2-oxoazetidin-3-ylhydroxymethyl)-pyrrolidine(3.34 g, 6.50 mmol) in tetrahydrofuran (35 ml) and the obtained mixturewas stirred at the same temperature for 25 minutes. To this solution wasadded a saturated aqueous solution of sodium chloride (35 ml) followedby extraction with diethyl ether (150 ml). The organic layer was washedsuccessively with water and a saturated aqueous solution of sodiumchloride. Next, the organic layer was dried over anhydrous magnesiumsulfate and concentrated under a reduced pressure. The residue wassubjected to silica gel chromatography [Wakogel C-200, 25 g,hexane-ethyl acetate (3:1 to 1:2)] to thereby give the target compound(2.30 g, 88.5%).

NMR(CDCl₃) δ: 0.05, 0.06(6H, each s), 0.86, 0.87(9H, each s), 1.47(9H,s), 1.70˜2.18(2H, m), 3.14˜3.46(4H, m), 3.48˜3.59(1H, m), 3.89˜4.10(1H,m), 4.20˜4.46(2H, m), 5.15(1H, d, J=8 Hz), 5.75(1H, br s)

Production Example 3

(2S,4R)-N-tert-Butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-ylhydroxymethyl)-pyrrolidine##STR28##

Under ice-cooling, a borane-methyl sulfide complex (2.3 ml, 23 mmol) wasadded to a solution of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethyl-siloxy-2-(2-oxoazetidin-3-ylhydroxymethyl)-pyrrolidine(1.84 g, 4.60 mmol) in tetra-hydrofuran (30 ml) and the obtained mixturewas stirred at the same temperature for 30 minutes and then heated underreflux for 8.5 hours. The reaction mixture was cooled with ice and thenmethanol (15 ml) was added thereto. After stirring at the sametemperature for 15 minutes, the solvent was distilled off under areduced pressure. Then, methanol (20 ml) was added to the residue andthe mixture was stirred at room temperature for 15 minutes followed bythe distillation off of the solvent under a reduced pressure. To thesolution obtained by dissolving the residue in tetrahydrofuran (20 ml)was added a solution of triethylamine (0.77 ml, 5.5 mmol) andp-nitro-benzyl chloroformate (1.09 g, 5.06 mmol) in tetrahydrofuran (5ml) under ice-cooling. The obtained mixture was stirred at the sametemperature for 3 hours. After adding ethyl acetate (150 ml) to thereaction mixture, the organic layer was washed successively with waterand a saturated aqueous solution of sodium chloride. Next, the organiclayer was dried over anhydrous magnesium sulfate and concentrated undera reduced pressure. The residue was subjected to silica gelchromatography [Wakogel C-200, 30 g, hexane-ethyl acetate (4:1 to 3:1)]to thereby give the target compound (450 mg, 17.3%).

NMR(CDCL₃) δ: 0.06(6H, s), 0.86(9H, s) 1.46, 1.47(9H, each s),1.58˜2.06(2H, m), 2.60˜2.76(1H, m), 3.08˜3.27(1H, m), 3.45˜3.59(1H, m),3.63˜3.73(1H, m), 3.74˜4.50(6H, m), 5.16(1H, d, J=19 Hz), 5.17(1H, d,J=19 Hz), 7.50, 7.55(2H, each d, J=8 Hz), 8.20, 8.23(2H, each d, J=8 Hz)

Production Example 4

(2S,4R)-4-Hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-ylhydroxymethyl)-pyrrolidine##STR29##

Under ice-cooling, trifluoroacetic acid (5 ml) was added to a solutionof(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyl-dimethylsiloxy-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-ylhydroxymethyl]-pyrrolidine(527 mg, 0.933 mmol) in methylene chloride (5 ml) and the obtainedmixture was stirred at the same temperature for 5 minutes and then atroom temperature for additional 50 minutes. After distilling off thesolvent and trifluoroacetic acid under a reduced pressure, the obtainedresidue was dissolved in tetrahydrofuran (7.5 ml). To this solution wereadded under ice-cooling triethylamine (0.53 ml, 3.8 mmol) andp-nitrobenzyl chloroformate (221 mg, 1.03 mmol) and the resultingmixture was stirred at the same temperature for 1 hour. After addingethyl acetate (60 ml) to the reaction mixture, the organic layer waswashed successively with water, 1 N hydrochloric acid, water, asaturated aqueous solution of sodium hydrogencarbonate and a saturatedaqueous solution of sodium chloride. Next, the organic layer was driedover anhydrous magnesium sulfate and concentrated under a reducedpressure. To the solution of the residue thus obtained intetrahydrofuran (6.5 ml) was added under ice-cooling a 1 M solution oftetrabutylammonium fluoride in tetrahydrofuran (1.9 ml, 1.9 mmol) andthe obtained mixture was stirred at the same temperature for 30 minutesand then at room temperature for additional 3.0 hours. After distillingoff the solvent under a reduced pressure, ethyl acetate (60 ml) wasadded to the residue. The organic layer was washed successively withwater, a 50%-saturated aqueous solution of sodium chloride and asaturated aqueous solution of sodium chloride. The organic layer wasdried over anhydrous magnesium sulfate and concentrated under a reducedpressure. The residue was subjected to silica gel chromatography(Wakogen C-200, 9 g, ethyl acetate) to thereby give the target compound(215 mg, 43.5%).

NMR(CDCl₃) δ: 1.75˜2.25(2H, m), 2.50˜2.76(1H, m), 3.44(1H, dd, J=3, 11Hz), 3.71(1H, d, J=12 Hz), 3.83˜4.31(6H, m), 4.46(1H, br s), 5.14(2H,s), 5.21(1H, d, J=19 Hz), 5.22(1H, d, J=19 Hz), 7.43˜7.53(4H, m),8.16(4H, d, J=8 Hz)

Production Example 5

(2S,4S)-4-Acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-ylhydroxymethyl)pyrrolidine##STR30##

Under ice-cooling, triethylamine (0.10 ml, 0.72 mmol) andmethanesulfonyl chloride (0.053 ml, 0.68 mmol) were added to a solutionof(2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-ylhydroxymethyl)-pyrrolidine(208 mg, 0.392 mmol) in methylene chloride (5 ml) and the obtainedmixture was stirred at the same temperature for 1 hour. Then thereaction mixture was washed successively with water, a saturated aqueoussolution of sodium hydrogen-carbonate and a saturated aqueous solutionof sodium chloride. The organic layer was dried over anhydrous magnesiumsulfate and concentrated under a reduced pressure. To the solution ofthe obtained residue in dimethylformamide (3 ml) was added potassiumthioacetate (150 mg, 1.18 mmol) and the mixture was stirred at 70° C.for 3 hours. After distilling off the solvent under a reduced pressure,ethyl acetate (40 ml) was added to the residue. The organic layer waswashed successively with water, a saturated aqueous solution of sodiumhydrogen-carbonate, water and a saturated aqueous solution of sodiumchloride. Next, the organic layer was dried over anhydrous magnesiumsulfate and concentrated under a reduced pressure. The residue wassubjected to silica gel chromatography [Wakogel C-200, 3 g, ethylacetate-hexane (1:1)] to thereby give the target compound (129 mg,56.0%).

NMR(CDCl₃) δ: 1.82˜1.96(1H, m), 2.30(1H, td, J=7, 13 Hz), 2.34(3H, s),2.63˜2.76(1H, m), 3.09(1H, t, J=11 Hz), 3.50˜4.09(6H, m), 4.13˜4.27(2H,m), 5.17(2H, s), 5.22(2H, s), 7.50(2H, d, J=8 Hz), 7.51(2H, d, J=8 Hz),8.18˜8.27(4H, m)

Example 1

p-Nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitro-benzyl-oxycarbonyl)azetidin-3-ylhydroxy-methyl)-pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylate##STR31##

Under ice-cooling, a 1 N aqueous solution of sodium hydroxide (0.28 ml,0.28 mmol) was added to a solution of(2S,4R)-4-acetylthio-N-(p-nitrobenzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-ylhydroxymethyl)-pyrrolidine(129 mg, 0.219 mmol) in a mixture of methanol (1.5 ml) withtetrahydrofuran (1.5 ml) and the obtained mixture was stirred at thesame temperature for 50 minutes. After adding 1 N hydrochloric acid(0.30 ml, 0.30 mmol) to the reaction mixture, the solvent was distilledoff under a reduced pressure and ethyl acetate (25 ml) was added to theresidue. Then, the organic layer was washed successively with water anda saturated aqueous solution of sodium chloride, dried over anhydrousmagnesium sulfate and concentrated under a reduced pressure. To thesolution of the obtained residue in acetonitrile (2 ml) were added,under ice-cooling, p-nitro-benzyl(1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxy-ethyl]-1-methylcarbapen-2-em-3-carboxylate(118 mg, 1.99 mmol) and diisopropyl-ethylamine (0.036 ml, 0.21 mmol).The obtained mixture was stirred at the same temperature for 10 minutesand then at room temperature for additional 1 hour. After adding ethylacetate (50 ml) to the reaction mixture, the organic layer was washedsuccessively with water, a 50%-saturated aqueous solution of sodiumchloride and a saturated aqueous solution of sodium chloride. Next, theorganic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography (Wakogel C-200, 3 g, 5% methanol-chloroform)to thereby give the target compound (137 mg, 70.3%).

NMR(CDCl₃) δ: 1.27(3H, d, J=7 Hz), 1.35(3H, d, J=7 Hz), 1.85˜2.40(2H,m), 2.52˜2.80(1H, m), 3.15˜3.26(1H, m), 3.28(1H, dd, J=2, 7 Hz),3.35(1H, dq, J=7, 7 Hz), 3.45˜3.65(1H, m), 3.83˜4.32(9H, m),5.10˜5.30(5H, m), 5.49(1H, d, J=13 Hz), 7.49(2H, d, J=8 Hz), 7.50(2H, d,J=8 Hz), 7.63(2H, d, J=8 Hz), 8.19(6H, d, J=8 Hz)

Example 2

(1R,5S,6S)-2-[(2S,4S)-2-(Azetidin-3-ylhydroxymethyl)-pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methylcarbapen-2-em-3-carboxylicacid acetate ##STR32##

To a solution of p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitro-benzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)-azetidin-3-ylhydroxymethyl)pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylate(137 mg, 0.154 mmol) in a mixture of a 0.1 M phosphate buffer (pH 7.0, 5ml) with tetrahydrofuran (5 ml) was added 20% palladium hydroxide/carbon(0.14 g) and hydrogenation was effected by using a Perlman shaker undera moderate pressure (4 kg/cm²) at room temperature for 2 hours. Afterfiltering off the catalyst through celite, the organic solvent wasdistilled off from the filtrate under a reduced pressure. The remainingaqueous layer was washed with diethyl ether and then the organic solventwas distilled off again from the aqueous layer under a reduced pressure.The pH value of the remaining aqueous layer was adjusted to 7.5 by usinga 1 N aqueous solution of sodium hydroxide. After filtering off theinsoluble matters, the filtrate was subjected to reversed phase silicagel chromatography (YMC SH-343-7 AM ODS, 0.005 M acetic acid-2.5%aqueous solution of methanol). The target fraction was concentrated andthen freeze-dried. The diastereomer mixture thus obtained was subjectedagain to reversed phase silica gel chromatography (YMC SH-343-7 AM ODS,0.02% ammonium acetate-5 to 20% aqueous solution of methanol) to therebyseparate the diastereomers from each other. The target fractions wereeach concentrated and subjected again to reversed phase silica gelchromatography (YMC SH-343-7 AM ODS, 0.005 M acetic acid-3% aqueoussolution of methanol) and each target fraction was concentrated andfreeze-dried. Thus, the diastereomer A (7.9 mg, 11.2%, the compound ofhigh polarity) and the diastereomer B (5.8 mg, 8.2%, the compound of lowpolarity) of the target compound were obtained.

Diastereomer A:

NMR(D₂ O) δ: 1.15(3H, d, J=7 Hz), 1.23(3H, d, J=6 Hz), 1.74(1H, td,J=10, 14 Hz), 1.87(3H, s), 2.72(1H, ddd, J=7, 7, 14 Hz), 2.97˜3.21(3H,m), 3.23˜3.37(2H, m), 3.42(1H, dd, J=2, 6 Hz), 3.52(2H, d, J=10 Hz),3.73˜3.83(1H, m), 4.07(1H, dd, J=6, 12 Hz), 4.15˜4.30(3H, m), 4.44(1H,d, J=4 Hz)

M.S. (m/e): 398(MH⁺)

Diastereomer B:

NMR(D₂ O) δ: 1.16(3H, d, J=7 Hz), 1.23(3H, d, J=6 Hz), 1.88(3H, s),2.16˜2.25(1H, m), 2.52˜2.65(1H, m), 2.72(1H, ddd, J=7, 7, 14 Hz),3.18(1H, dd, J=7, 13 Hz), 3.24˜3.39(4H, m), 3.46(1H, dd, J=2, 6 Hz),3.77(1H, dd, J=6, 13 Hz), 3.98˜4.14(3H, m), 4.15˜4.25(2H, m), 4.43(1H,dd, J=8, 9 Hz)

MS (m/e): 398(MH⁺)

Production Example 6

(2S,4R)-N-tert-Butoxycarbonyl-2-(N-tert-butoxycarbonyl-2-oxypyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethyl-siloxy-pyrrolidine##STR33##

Under ice-cooling, a 1.7 M solution of n-butyl-lithium in hexane (113ml, 192 mmol) was added to a solution of hexa-methyldisilazane (42.3 ml,200 mmol) in tetrahydrofuran (300 ml). After stirring at the sametemperature for 25 minutes, the reaction mixture was cooled to -78° C.Into this solution was dropped a solution ofN-tert-butoxycarbonyl-2-oxo-pyrrolidine (337 g, 182 mmol) intetrahydrofuran (150 ml) and the resulting mixture was stirred at -78°C. for 45 minutes. This solution was dropped into a solution of(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-formylpyrrolidine(60.0 g, 182 mmol) in tetrahydrofuran (500 ml) cooled to -78° C. andstirred at the same temperature for 25 minutes. After adding a saturatedaqueous solution of ammonium chloride (120 ml), the reaction mixture wasreturned to room temperature. Then the tetrahydrofuran was distilled offunder a reduced pressure and ethyl acetate (900 ml) was added to theresidue. The organic layer was washed successively with water, 1 Nhydrochloric acid, water, a saturated aqueous solution of sodiumhydrogencarbonate and a saturated aqueous solution of sodium chloride.The organic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography [Merck, kieselgel 60, 1.4 kg, hexane-ethylacetate (9:1 to 7:3)] to thereby give the diastereomer A (8.22 g, 8.8%,the compound of low polarity), the diastereomer B (28.3 g, 30.2%, thecompound of moderate polarity) and the diastereomer C (24.1 g, 25.7%,the compound of high polarity) of the target compound.

Diastereomer A:

NMR(CDCl₃) δ: 0.06(6H, s), 0.88(9H, s), 1.45, 1.53(18H, each s),1.80˜2.13(3H, m), 2.40˜2.50(1H, m), 2.55(1H, dd, J=9, 16 Hz), 3.31(1H,dd, J=4, 11 Hz), 3.48˜3.68(3H, m), 3.77˜3.85(1H, m), 4.32˜4.43(2H, m),5.62(1H, br s)

Diastereomer B:

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.45, 1.53(18H, each s),1.64˜1.70(1H, m), 1.83˜2.13(2H, m), 2.24(1H, td, J=5, 13 Hz), 2.40(1H,dd, J=11, 20 Hz), 3.28˜3.62(3H, m), 3.77˜3.98(2H, m), 4.13˜4.54(3H, m)

Diastereomer C:

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.45, 1.53(18H, each s),1.82˜1.92 (1H, m), 1. 94˜2.05 (2H, m), 2.15˜2.27(1H, m), 2.54(1H, t,J=10 Hz), 3.23(1H, dd, J=4, 11 Hz), 3.51(1H, d, J=12 Hz), 3.56(1H, td,J=8, 11 Hz), 3.74˜3.82(1H, m), 4.18˜4.48(3H, m)

Production Example 7

(2S,4R) -N-tert-Butoxycarbonyl-2-(N-tert-butoxycarbonyl-pyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer A ##STR34##

At room temperature, a borane-methyl sulfide complex (0.64 ml, 6.4 mmol)was added to a solution of(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxy-carbonyl-2-oxopyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxypyrrolidinediastereomer A (1.10 g, 2.14 mmol) in tetrahydrofuran (15 ml) and theobtained mixture was heated under reflux for 1.5 hours. Afterice-cooling the reaction mixture, methanol (3 ml) was added thereto andthe obtained mixture was stirred at the same temperature for 10 minutes.Then, the solvent was distilled off under a reduced pressure and theresidue was subjected to silica gel chromatography [Wakogel C-200, 15 g,hexane-ethyl acetate (5:1)] to thereby give the target compound (825 mg,77.1%).

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.46(18H, s), 1.53˜1.73(1H, m),1.76˜1.89(1H, m), 1.91˜2.23(3H, m), 3.14˜3.27(2H, m), 3.30˜3.75(5H, m),3.96(1H, q, J=8 Hz), 4.08˜4.32(1H, m), 5.36, 5.44(1H, each brs)

Production Example 8

(2S,4R)-4-Hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer A ##STR35##

Under ice-cooling, trifluoroacetic acid (5 ml) was added to a solutionof(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonylpyrrolidin-3-ylhydroxymethyl)-4-tert-butyldi-methylsiloxy-pyrrolidinediasteromer A (815 mg, 1.63 mmol) in methylene chloride (5 ml) and theobtained mixture was stirred at the same temperature for 10 minutes andthen at room temperature for an additional 1.5 hours. After distillingoff the solvent and trifluoroacetic acid under a reduced pressure, theobtained residue was dissolved in tetrahydrofuran (13 ml). To thissolution was added, under ice-cooling, a solution of triethylamine (1.4ml, 10 mmol) and p-nitrobenzyl chloro-formate (0.74 g, 3.4 mmol) intetrahydrofuran (4 ml) and the resulting mixture was stirred at the sametemperature for 1 hour. After distilling off the solvent under a reducedpressure, ethyl acetate (40 ml) was added to the residue. Then, theorganic layer was washed successively with water, 1 N hydrochloric acid,water, a saturated aqueous solution of sodium hydrogencarbonate and asaturated aqueous solution of sodium chloride. Next, the organic layerwas dried over anhydrous magnesium sulfate and concentrated under areduced pressure. To the solution of the residue thus obtained intetrahydrofuran (15 ml) was added, under ice-cooling, a 1 M solution oftetrabutylammonium fluoride in tetrahydrofuran (2.25 ml, 2.25 mmol) andthe obtained mixture was stirred at the same temperature for 10 minutesand then at room temperature for an additional 2.5 hours. Afterdistilling off the solvent under a reduced pressure, ethyl acetate (40ml) was added to the residue. The organic layer was washed successivelywith water, a 50%-saturated aqueous solution of sodium chloride and asaturated aqueous solution of sodium chloride. The organic layer wasdried over anhydrous magnesium sulfate and concentrated under a reducedpressure. The residue was subjected to silica gel chromatography(Wakogel C-200, 8 g, ethyl acetate) to thereby give the target compound(393 mg, 44.3%).

NMR(CDCl₃) δ: 1.71˜2.18(5H, m), 2.19˜2.35(1H, m), 3.24˜3.72(6H, m),3.78˜3.91(1H, m), 4.05˜4.31(1H, m), 4.42˜4.50(1H, m), 4.81˜4.98(1H, m),5.22(2H, s), 5.25, 5.26(2H, each s), 7.48˜7.55(4H, m), 8.19˜8.25(4H, m)

Production Example 9

(2S,4S)-4-Acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl-hydroxymethyl]-pyrrolidinediastereomer A ##STR36##

Under ice-cooling, triethylamine (0.13 ml, 0.93 mmol) andmethanesulfonyl chloride (0.07 ml, 0.90 mmol) were added to a solutionof(2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-nitrobenzyloxy-carbonyl)-pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer A (384 mg, 0.71 mmol) in methylene chloride (5 ml) and theobtained mixture was stirred at the same temperature for 30 minutes.Then, the reaction mixture was washed successively with water, asaturated aqueous solution of sodium hydrogencarbonate and a saturatedaqueous solution of sodium chloride. The organic layer was dried overanhydrous magnesium sulfate and concentrated under a reduced pressure.To the solution of the obtained residue in dimethyl-formamide (5 ml) wasadded potassium thioacetate (242 mg, 2.12 mmol) and the mixture wasstirred at 70° C. for 3.5 hours. After distilling off the solvent undera reduced pressure, ethyl acetate (40 ml) was added to the residue. Theorganic layer was washed successively with water, a saturated aqueoussolution of sodium hydrogencarbonate, water and a saturated aqueoussolution of sodium chloride. Next, the organic layer was dried overanhydrous magnesium sulfate and concentrated under a reduced pressure.The residue was subjected to silica gel chromatography [Wakogen C-200, 5g, ethyl acetate-hexane (1:1)] to thereby give the target compound (203mg, 47.8%).

NMR(CDCl₃) δ: 1.67˜1.77(1H, m), 1.85˜2.13(2H, m), 2.20˜2.47(1H, m),2.35, 2.36(3H, each s), 2.47˜2.61 (1H, m), 3.17(1H, ddd, J=2, 10, 12Hz), 3.26˜3.54(3H, m), 3.56˜3.75(2H, m), 3.80(1H, ddd, J=7, 9, 15 Hz),3.91˜4.03(1H, m), 4.19˜4.27(1H, m), 4.69˜4.81(1H, m), 5.18˜5.29(4H, m),7.52(4H, d, J=8 Hz), 8.19˜8.27(4H, m)

Example 3

p-Nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitro-benzyloxy-carbonyl)-pyrrolidin-3-ylhydroxymethyl]-pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylatediastereomer A ##STR37##

Under ice-cooling, a 1 N aqueous solution of sodium hydroxide (0.34 ml,0.34 mmol) was added to a solution of (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer A (196 mg, 0.33 mmol) in a mixture of methanol (3 ml) withtetrahydrofuran (3 ml) and the obtained mixture was stirred at the sametemperature for 20 minutes. After adding 1 N hydrochloric acid (0.36 ml,0.36 mmol) to the reaction mixture, the solvent was distilled off undera reduced pressure and ethyl acetate (30 ml) was added to the residue.Then, the organic layer was washed successively with water and asaturated aqueous solution of sodium chloride, dried over anhydrousmagnesium sulfate and concentrated under a reduced pressure. To thesolution of the obtained residue in acetonitrile (4 ml) were added,under ice-cooling, p-nitro-benzyl(1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methylcarbapen-2-em-3-carboxylate(177 mg, 0.30 mmol) and diisopropyl-ethylamine (0.055 ml, 0.32 mmol).The obtained mixture was stirred at the same temperature for 40 minutesand then at room temperature for an additional 1.5 hours. After addingethyl acetate (40 ml) to the reaction mixture, the organic layer waswashed successively with water, a 50%-saturated aqueous solution ofsodium chloride and a saturated aqueous solution of sodium chloride.Next, the organic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography (Wakogel C-200, 5 g, 1% methanol-chloroform)to thereby give the target compound (204 mg, 73.0%).

NMR(CDCl₃) δ: 1.28(3H, d, J=7 Hz), 1.38(3H, d, J=6 Hz), 1.69˜1.82(1H,m), 1.79(1H, d, J=4 Hz), 1.88˜2.15(2H, m), 2.20˜2.40(1H, m),2.41˜2.64(1H, m), 3.21˜3.80(9H, m), 3.92˜4.07(1H, m), 4.07˜4.21(1H, m),4.23˜4.30(2H, m), 5.15˜5.30(5H, m), 5.50(1H, d, J=13 Hz), 7.52(4H, d,J=8 Hz), 7.65(2H, d, J=9 Hz), 8.18˜8.27(6H, m)

Example 4

(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-3-ylhydroxymethyl)pyrrolidin-4-ylthio]-carbapen-2-em-3-carboxylicacid acetate diastereomer A ##STR38##

To a solution of p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxy-carbonyl)pyrrolidin-3-ylhydroxymethyl]pyrolidin-4-ylthio}carbapen-2-em-3-carboxylatediastereomer A (204 mg, 0.127 mmol) in a mixture of a 0.1 M phosphatebuffer (pH 7.0, 5 ml) with tetrahydrofuran (5 ml) was added 20%palladium hydroxide/carbon (205 mg) and hydrogenation was effected byusing a Perlman shaker under a moderate pressure (4 kg/cm²) at roomtemperature for 2 hours. After filtering off the catalyst throughcelite, the organic solvent was distilled off from the filtrate under areduced pressure. The remaining aqueous layer was washed with diethylether and then the organic solvent was distilled off again from theaqueous layer under a reduced pressure. The pH value of the remainingaqueous layer was adjusted to 7.5 by using a 1 N aqueous solution ofsodium hydroxide. After filtering off the insoluble matters, thefiltrate was subjected to reversed phase silica gel chromatography (YMCSH-343-7 AM ODS, 0.005 M acetic acid-3% aqueous solution of methanol).The target fraction was concentrated and then freeze-dried to therebygive the target compound (41.7 mg, 40.7%).

NMR(D₂ O) δ: 1.16(3H, d, J=7 Hz), 1.23(3H, d, J=6 Hz), 1.68˜1.89(2H, m),1.86(3H, s), 2.10˜2.23(1H, m), 2.46˜2.58(1H, m), 2.65(1H, td, J=7, 16Hz), 3.13˜3.46(7H, m), 3.60(1H, dd, J=7, 12 Hz), 3.67(1H, q, J=8 Hz),3.92(1H, dd, J=6, 7 Hz), 3.94˜4.03(1H, m), 4.14˜4.24(2H, m)

M.S. (m/e): 412(MH⁺)

Production Example 10

(2S,4R)-N-tert-Butoxycarbonyl-2-(N-tert-butoxycarbonyl-pyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer B ##STR39##

The reaction of Production Example 7 was repeated by using(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonyl-2-oxopyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethyl-siloxypyrrolidinediastereomer B (4.51 g, 8.77 mmol) and the borane-methyl sulfide complex(2.6 ml, 26 mmol) to thereby give the target compound (3.52 g, 80.3%).

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.45, 1.46(18H, each s),1.65˜2.20(5H, m), 3.08˜3.73(6H, m), 3.90˜4.20(2H, m), 4.24˜4.38(1H, m)

Production Example 11

(2S,4R)-4-Hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer B ##STR40##

The reaction of Production Example 8 was repeated by using (2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonyl-pyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer B (17.2 mg, 34.4 mmol) to thereby give the target compound(11.4 g, 61.0%).

NMR(CDCl₃) δ: 1.71˜-2.00(4H, m), 2.09˜2.39(2H, m), 3.27 (1H, t, J=8 Hz),3.36 (1H, td, J=7, 10 Hz), 3.48(1H, dd, J=3, 10 Hz), 3.56˜3.88(3H, m),4.12˜4.25(2H, m), 4.48˜4.54(1H, brs), 5.13˜5.29(4H, m), 7.50(4H, d, J=8Hz), 8.19(2H, d, J=8 Hz), 8.20(2H, d, J=8 Hz)

Production Example 12

(2S,4S)-4-Acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl-hydroxymethyl]-pyrrolidinediastereomer B ##STR41##

The reaction of Production Example 9 was repeated by using(2S,4R)-4-hydroxy-N-(p-nitrobenzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer B (10.3 g, 19.0 mmol) to thereby give the target compound(5.93 g, 51.9%).

NMR(CDCl₃) δ: 1.82˜2.42(5H, m), 2.35(3H, s), 3.05˜3.17(1H, m),3.21˜3.32(1H, m), 3.35(1H, dt, J=7, 10 Hz), 3.52˜3.88(3H, m),3.93˜4.27(3H, m), 5.21(4H, s), 7.51(4H, d, J=8 Hz), 8.17˜8.28(4H, m)

Example 5

p-Nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitro-benzyloxy-carbonyl)-pyrrolidin-3-ylhydroxymethyl]-pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylatediastereomer B ##STR42##

The reaction of Example 3 was repeated by using(2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyl-oxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer B (5.00 g, 8.30 mmol) to thereby give the target compound(6.40 g, 85.3%).

NMR(CDCl₃) δ: 1.28(3H, d, J=7 Hz), 1.37(3H, d, J=6 Hz), 1.50˜2.53(5H,m), 3.15˜3.45(5H, m), 3.47˜3.82(3H, m), 3.90˜4.32(5H, m), 5.14˜5.29(5H,m), 5.50(1H, d, J=13 Hz), 7.45˜7.55(4H, m), 7.65(2H, d, J=9 Hz),8.17˜8.26(6H, m)

Example 6

(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-3-ylhydroxymethyl)pyrrolidin-4-ylthio]carbapen-2-em-3-carboxylicacid acetate diastereomer B ##STR43##

The reaction of Example 4 was repeated by using p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyl-oxycarbonyl)pyrrolidin-3-ylhydroxymethyl]pyrrolidin-4-ylthio}carbapen-2-em-3-carboxylatediastereomer B (6.37 g, 7.05 mmol) to thereby give the target compound(1.25 g, 37.7%).

NMR(D₂ O) δ: 1.16(3H, d, J=7 Hz), 1.23(3H, d, J=6 Hz), 1.65˜1.78(1H, m),1.78˜1.90(1H, m), 1.85(3H, s), 2.07˜2.18(1H, m), 2.43(1H, qd, J=9, 18Hz), 2.57(1H, td, J=8, 14 Hz), 3.15(1H, dd, J=10, 12 Hz), 3.20˜3.36(3H,m), 3.38˜3.47(2H, m), 3.50(1H, dd, J=8, 12 Hz), 3.63(1H, dd, J=7, 12Hz), 3.80(1H, ddd, J=4, 8, 12 Hz), 3.90˜4.00(2H, m), 4.14˜4.23(2H, m)

M.S. (m/e): 412(MH⁺)

Production Example 13

(2S,4R)-N-tert-Butoxycarbonyl-2-(N-tert-butoxycarbonyl-pyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer C ##STR44##

The reaction of Production Example 7 was repeated by using(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonyl-2-oxopyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethyl-siloxypyrrolidinediastereomer C (24.1 g, 46.8 mmol) and the borane-methyl sulfide complex(25.3 ml, 253 mmol) to thereby give the target compound (20.2 g, 86.3%).

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.46(18H, s), 1.56˜2.24(5H, m),2.85˜3.32(3H, m), 3.34˜3.71(3H, m), 3.80˜4.20(2H, m), 4.24˜4.42(1H, m)

Production Example 14

(2S,4R)-4-Hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer C ##STR45##

The reaction of Production Example 8 was repeated by using(2S,4S)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonyl-pyrrolidin-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer C (2.93 mg, 5.86 mmol) to thereby give the target compound(1.65 g, 51.8%).

NMR(CDCl₃) δ: 1.67˜1.87(2H, m), 1.89˜2.01(1H, m), 2.03˜2.37(3H, m),3.10˜3.86(6H, m), 3.94˜4.20(2H, m), 4.46˜4.55(1H, m), 5.15˜5.29(4H, m),7.51(2H, d, J=8 Hz), 8.21(2H, d, J=8 Hz), 8.22(2H, d, J=8 Hz)

Production Example 15

(2S,4S)-4-Acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl-hydroxymethyl]-pyrrolidinediastereomer C ##STR46##

The reaction of Production Example 9 was repeated by using(2S,4R)-4-hydroxy-N-(p-nitrobenzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer C (7.93 g, 14.6 mmol) to thereby give the target compound(4.57 g, 52.0%).

NMR(CDCl₃) δ: 1.67˜1.88(1H, m), 1.94˜2.24(3H, m), 2.29˜2.48(1H, m),2.33, 2.34(3H, each s), 3.05˜3.25(2H, m), 3.27˜3.39(1H, m),3.48˜3.67(2H, m), 3.74˜3.86(1H, m), 3.88˜3.98(1H, m), 4.05˜4.27(2H, m),5.21(4H, s), 7.51(4H, d, J=8 Hz), 8.19˜8.27(4H, m)

Example 7

p-Nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxy-carbonyl)-pyrrolidin-3-ylhydroxymethyl]pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylatediastereomer C ##STR47##

The reaction of Example 3 was repeated by using(2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyl-oxycarbonyl)pyrrolidin-3-ylhydroxymethyl]-pyrrolidinediastereomer C (4.00 g, 6.64 mmol) to thereby give the target compound(5.04 g, 84.0%).

NMR(CDCl₃) δ: 1.28(3H, d, J=7 Hz), 1.38(3H, d, J=6 Hz), 1.50˜2.50(5H,m), 3.08˜3.43(5H, m), 3.46˜3.69(3H, m), 3.84˜4.31(5H, m), 5.15˜5.29(5H,m), 5.50(1H, d, J=13 Hz), 7.44˜7.57(4H, m), 7.65(2H, d, J=9 Hz),8.17˜8.27(6H, m)

Example 8

(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-3-ylhydroxymethyl)pyrrolidin-4-ylthio]carbapen-2-em-3-carboxylicacid acetate diastereomer C ##STR48##

The reaction of Example 4 was repeated by using p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxy-carbonyl)pyrrolidin-3-ylhydroxymethyl]pyrrolidin-4-ylthio}carbapen-2-em-3-carboxylatediastereomer C (4.99 g, 5.50 mmol) to thereby give the target compound(856 mg, 33.0%).

NMR(D₂ O) δ: 1.16(3H, d, J=7 Hz), 1.23(3H, d, J=6 Hz), 1.81˜1.94(2H, m),1.87(3H, s), 2.13˜2.23(1H, m), 2.45(1H, qd, J=8, 16 Hz), 2.60(1H, td,J=8, 14 Hz), 3.01(1H, dd, J=9, 11 Hz), 3.20˜3.35(3H, m), 3.36˜3.47(3H,m), 3.63(1H, dd, J=7, 12 Hz), 3.75(1H, ddd, J=4, 9, 10 Hz),3.92˜4.00(2H, m), 4.15˜4.24(2H, m)

M.S. (m/e): 412 (MH⁺)

Example 9

(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-3-ylhydroxymethyl)pyrrolidin-4-ylthio]carbapen-2-em-3-carboxylicacid hydrochloride diastereomer B ##STR49##

To a solution of p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitro-benzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)-pyrrolidin-3-ylhydroxymethyl}pyrrolidin-4-ylthio}carbapen-2-em-3-carboxylatediastereomer B (1.05 g, 1.16 mmol) in a mixture of a 0.1 M phosphatebuffer (pH 7.0, 15 ml) with tetrahydrofuran (15 ml) was added 20%palladium hydroxide/carbon (1.05 g) and hydrogenation was effected byusing a Perlman shaker under a moderate pressure (4 kg/cm²) at roomtemperature for 2 hours. After filtering off the catalyst throughcelite, the organic solvent was distilled off from the filtrate under areduced pressure. The remaining aqueous layer was washed with diethylether and then the organic solvent was distilled off again from theaqueous layer under a reduced pressure. After filtering off theinsoluble matters, the pH value of the remaining aqueous layer wasadjusted to 7.5 by using a 1 N aqueous solution of sodium hydroxide.Then, the filtrate was subjected to reversed phase silica gelchromatography (YMC SH-343-7 AM ODS, water -20% aqueous solution ofmethanol). The target fraction was concentrated and the pH value thereofwas adjusted to 6.0 with 1 N hydrochloric acid. After freeze-drying, thetarget compound (147 mg, 28.3%) was obtained.

NMR(D₂ O) δ: 1.17(3H, d, J=7 Hz), 1.24(3H, d, J=6 Hz), 1.73(1H, qd, J=9,13 Hz), 1.84(1H, ddd, J=7, 10, 12 Hz), 2.07˜2.18(1H, m), 2.44(1H, qd,J=9, 18 Hz), 2.58(1H, td, J=8, 14 Hz), 3.15(1H, dd, J=10, 12 Hz), 3.213.37(3H, m) 3.39˜3.47(2H, m), 3.51(1H, dd, J=8, 12 Hz), 3.64(1H, dd,J=7, 12 Hz), 3.83(1H, ddd, J=3, 8, 11 Hz), 3.92˜4.01(2H, m),4.15˜4.23(2H, m)

Production Example 16

(2S,4R)-N-tert-Butoxycarbonyl-2-(N-tert-butoxycarbonyl-2-oxopiperidin-3-ylhydroxymethyl)-4-tert-butyldimethyl-siloxy-pyrrolidine##STR50##

Under ice-cooling, a 2.5 M solution of n-butyl-lithium in hexane (12.6ml, 31.5 mmol) was added to a solution of hexa-methyldisilazane (7.0 ml,33 mmol) in tetrahydrofuran (50 ml). After stirring at the sametemperature for 20 minutes, the reaction mixture was cooled to -78° C.Into this solution was dropped a solution ofN-tert-butoxycarbonyl-2-oxopiperidine (5.97 g, 30.0 mmol) intetrahydrofuran (30 ml) and the resulting mixture was stirred at -78° C.for 50 minutes. This solution was dropped into a solution of(2S,4R)-N-tert-butoxy-carbonyl-4-tert-butyldimethylsiloxy-2-formylpyrrolidine(9.87 g, 30.0 mmol) in tetrahydrofuran (100 ml) cooled to -78° C. andstirred at the same temperature for 40 minutes. After adding a saturatedaqueous solution of ammonium chloride (20 ml), the reaction mixture wasreturned to room temperature. Then, the tetrahydrofuran was distilledoff under a reduced pressure and ethyl acetate (220 ml) was added to theresidue. The organic layer was washed successively with water, 1 Nhydrochloric acid, water, a saturated aqueous solution of sodiumhydrogen-carbonate and a saturated aqueous solution of sodium chloride.The organic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography [Wakogel C-200, 150 g, hexane-ethyl acetate(9:1 to 3:1)] to thereby give the diastereomer A (5.50 g, 31.5%, thecompound of low polarity) and the diastereomer B (3.62 g, 20.8%, thecompound of high polarity) of the target compound were obtained.

Diastereomer A:

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.44(9H, s), 1.51(9H, s),1.67˜2.00(4H, m), 2.17˜2.44(2H, m), 3.29˜3.80(5H, m), 3.88˜4.33(3H, m),4.45˜4.53(2H, m)

Diastereomer B:

NMR(CDCl₃) δ: 0.06(6H, S), 0.86(9H, S), 1.45(9H, s), 1.52(9H, s)1.77˜1.96(3H, m), 2.09˜2.26(2H, m), 2.50˜2.63(1H, m), 3.27(1H, dd, J=4,11 Hz), 3.37˜3.64(3H, m), 3.74˜3.84(1H, m), 3.99˜4.45(4H, m)

Production Example 17

(2S,4R)-N-tert-Butoxycarbonyl-2-(N-tert-butoxycarbonyl-piperidine-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer A ##STR51##

At room temperature, a borane-methyl sulfide complex (5.60 ml, 56.0mmol) was added to a solution of(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonyl-2-oxopiperidin-3-ylhydroxymethyl)-4-tert-butyldimethyl-siloxypyrrolidinediastereomer A (5.50 g, 10.4 mmol) in tetrahydrofuran (100 ml) and theobtained mixture was heated under reflux for 1.2 hours. Afterice-cooling the reaction mixture, methanol (26 ml) was added thereto andthe obtained mixture was stirred at the same temperature for 15 minutes.Then, the solvent was distilled off under a reduced pressure and theresidue was subjected to silica gel chromatography [Wakogel C-200, 70 g,hexane-ethyl acetate (9:1 to 5:1)] to thereby give the target compound(2.99 g, 55.9%).

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.45(18H, brs), 1.24˜2.18(7H,m), 3.01˜4.46(9H, m)

Production Example 18

(2S,4R)-4-Hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-3-ylhydroxymethyl]-pyrrolidinediastereomer A ##STR52##

Under ice-cooling, trifluoroacetic acid (15 ml) was added to a solutionof(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonylpiperidin-3-ylhydroxymethyl)-4-tert-butyldi-methylsiloxy-pyrrolidinediastereomer A (2.94 g, 5.72 mmol) in methylene chloride (15 ml) and theobtained mixture was stirred at the same temperature for 10 minutes andthen at room temperature for an additional 1.1 hours. After distillingoff the solvent and trifluoroacetic acid under a reduced pressure, theobtained residue was dissolved in tetrahydro-furan (55 ml). To thissolution was added, under ice-cooling, a solution of triethylamine (6.1ml, 44 mmol) and p-nitrobenzyl chloroformate (2.59 g, 12.0 mmol) intetrahydrofuran (15 ml) and the resulting mixture was stirred at thesame temperature for 40 minutes. After distilling off the solvent undera reduced pressure, ethyl acetate (100 ml) was added to the residue.Then, the organic layer was washed successively with water, 1 Nhydrochloric acid, water, a saturated aqueous solution of sodiumhydrogencarbonate and a saturated aqueous solution of sodium chloride.Next, the organic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. To the solution of the residuethus obtained in tetrahydrofuran (40 ml) was added, under ice-cooling, a1 M solution of tetrabutylammonium fluoride in tetrahydrofuran (11.5 ml,11.5 mmol) and the obtained mixture was stirred at the same temperaturefor 20 minutes and then at room temperature for an additional 4.0 hours.After distilling off the solvent under a reduced pressure, ethyl acetate(100 ml) was added to the residue. The organic layer was washedsuccessively with water, a 50%-saturated aqueous solution of sodiumchloride and a saturated aqueous solution of sodium chloride. Theorganic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography (Wakogel C-200, 40 g, ethyl acetate) tothereby give the target compound (1.47 g, 46.1%).

NMR(CDCl₃) δ: 1.20˜2.03(6H, m), 2.10˜2.33(1H, m), 2.81˜3.13(1H, m),3.33˜3.59(3H, m), 3.61˜3.86(2H, m), 3.89˜4.35(2H, m), 4.38˜4.59(1H, m),5.10˜5.33(4H, m), 7.41˜7.56(4H, m), 8.15˜8.26(4H, m)

Production Example 19

(2S,4S)-4-Acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-3-yl-hydroxymethyl]-pyrrolidinediastereomer A ##STR53##

Under ice-cooling, triethylamine (0.46 ml, 3.3 mmol) and methanesulfonylchloride (0.25 ml, 3.2 mmol) were added to a solution of(2S,4R)-4-hydroxy-N-(p-nitro-benzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)-piperidine-3-ylhydroxymethyl]-pyrrolidinediastereomer A (1.43 g, 2.56 mmol) in methylene chloride (15 ml) and theobtained mixture was stirred at the same temperature for 30 minutes.Then, the reaction mixture was washed successively with water, asaturated aqueous solution of sodium hydrogencarbonate and a saturatedaqueous solution of sodium chloride. The organic layer was dried overanhydrous magnesium sulfate and concentrated under a reduced pressure.To the solution of the obtained residue in dimethylformamide (15 ml) wasadded potassium thioacetate (0.98 mg, 7.7 mmol) and the mixture wasstirred at 70° C. for 5.5 hours. After distilling off the solvent undera reduced pressure, ethyl acetate (40 ml) was added to the residue. Theorganic layer was washed successively with water, a saturated aqueoussolution of sodium hydrogencarbonate, water and a saturated aqueoussolution of sodium chloride. Next, the organic layer was dried overanhydrous magnesium sulfate and concentrated under a reduced pressure.The residue was subjected to silica gel chromatography (wakogel C-200,15 g, ethyl acetate-hexane (1:1)] to thereby give the target compound(890 mg, 56.4%).

NMR(CDCl₃) δ: 1.20˜1.63(3H, m), 1.66˜1.82(2H, m), 1.87˜2.14(1H, m),2.20˜2.38(1H, m), 2.35(3H, s), 2.65˜4.32(9H, m), 5.08˜5.32(4H, m),7.40˜7.57(4H, m), 8.21(4H, d, J=9 Hz)

Example 10

p-Nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitro-benzyloxy-carbonyl)piperidine-3-ylhydroxymethyl]-pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylatediastereomer A ##STR54##

Under ice-cooling, a 1 N aqueous solution of sodium hydroxide (1.48 ml,1.48 mmol) was added to a solution of(2S,4S)-4-acetylthio-N-(p-nitrobenzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-3-ylhydroxymethyl]-pyrrolidinediastereomer A (869 mg, 1.41 mmol) in a mixture of methanol (8.5 ml)with tetrahydrofuran (8.5 ml) and the obtained mixture was stirred atthe same temperature for 30 minutes. After adding 1 N hydrochloric acid(1.55 ml, 1.55 mmol) to the reaction mixture, the solvent was distilledoff under a reduced pressure and ethyl acetate (60 ml) was added to theresidue. Then, the organic layer was washed successively with water anda saturated aqueous solution of sodium chloride, dried over anhydrousmagnesium sulfate and concentrated under a reduced pressure. To thesolution of the obtained residue in acetonitrile (10 ml) were added,under ice-cooling, p-nitrobenzyl(1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methylcarbapen-2-em-3-carboxylate(829 mg, 1. 40 mmol) and diisopropyl-ethylamine (0.26 ml, 1.5 mmol). Theobtained mixture was stirred at the same temperature for 10 minutes andthen at room temperature for an additional 4.5 hours. After adding ethylacetate (70 ml) to the reaction mixture, the organic layer was washedsuccessively with water, a 50%-saturated aqueous solution of sodiumchloride and a saturated aqueous solution of sodium chloride. Next, theorganic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography (Wakogel C-200, 23 g, 2% methanol-chloroform)to thereby give the target compound (1.04 g, 80.3%).

NMR(CDCl₃) δ: 1.15˜2.35(7H, m), 1.28(3H, d, J=7 Hz), 1.37(3H, d, J=6Hz), 2.85˜4.35(12H, m), 3.27(1H, dd, J=2.7 Hz), 5.10˜5.35(5H, m),5.50(1H, d, J=13 Hz), 7.40˜7.60(4H, m), 7.65(2H, d, J=9 Hz), 8.21(6H, d,J=9 Hz)

Example 11

(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-1-methyl-2-[(2S,4S)-2-piperidine-3-ylhydroxymethyl)pyrrolidin-4-ylthio]-carbapen-2-em-3-carboxylicacid acetate diastereomer A ##STR55##

To a solution of p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitro-benzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxy-carbonyl)piperidine-3-ylhydroxymethyl]pyrrolidin-4-ylthio}carbapen-2-em-3-carboxylatediastereomer A (1.00 g, 1.09 mmol) in a mixture of a 0.1 M phosphatebuffer (pH 7.0, 15 ml) with tetrahydrofuran (15 ml) was added 20%palladium hydroxide/carbon (1.0 g) and hydrogenation was effected byusing a Perlman shaker under moderate pressure (4 kg/cm²) at roomtemperature for 2 hours. After filtering off the catalyst throughcelite, the organic solvent was distilled off from the filtrate under areduced pressure. The remaining aqueous layer was washed with diethylether and then the organic solvent was distilled off again from theaqueous layer under a reduced pressure. The pH value of the remainingaqueous layer was adjusted to 7.5 by using a 1 N aqueous solution ofsodium hydroxide. After filtering off the insoluble matters, thefiltrate was subjected to reversed phase silica gel chromatography (YMCSH-343-7 AM ODS, 0.005 M acetic acid-4% aqueous solution of methanol).The target fraction was concentrated and then freeze-dried to therebygive the target compound (193 mg, 36.4%).

NMR(D₂ O) δ: 1.18(3H, d, J=7 Hz), 1.24(3H, d, J=6 Hz), 1.25˜1.37(1H, m),1.58˜1.73(1H, m), 1.73˜1.89(3H, m), 1.87(3H, s), 1.89˜1.98(1H, m),2.58(1H, td, J=8, 14 Hz), 2.77(1H, t, J=12 Hz), 2.87(1H, dd, J=3, 13Hz), 3.27˜3.40(3H, m), 3.42(1H, dd, J=2, 6 Hz), 3.53˜3.60(1H, m),3.65(1H, dd, J=7, 12 Hz), 3.80(1H, dd, J=3, 9 Hz), 3.90˜4.01(2H, m),4.16˜4.24(2H, m)

M.S. (m/e): 426(MH⁺)

Production Example 20

(2S,4R)-N-tert-Butoxycarbonyl-2-(N-tert-butoxycarbonyl-piperidine-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer B ##STR56##

The reaction of Production Example 17 was repeated by using(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonyl-2-oxopiperidin-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer B (3.60 g, 6.82 mmol) and the borane methyl sulfide complex(2.05 ml, 20.5 mmol) to thereby give the target compound (2.50 9,71.3%).

NMR(CDCl₃) δ: 0.06(6H, s), 0.86(9H, s), 1.46(18H, br s), 1.15˜2.26(7H,m), 2.46˜2.75(1H, m), 2.97˜4.44(8H, m)

Production Example 21

(2S,4R)-4-Hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-3-ylhydroxymethyl]-pyrrolidinediastereomer B ##STR57##

The reaction of Production Example 18 was repeated by using(2S,4R)-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonyl-piperidine-3-ylhydroxymethyl)-4-tert-butyldimethylsiloxy-pyrrolidinediastereomer B (2.48 mg, 4.82 mmol) to thereby give the target compound(778 mg, 28.9%).

NMR(CDCl₃) δ: 1.15˜2.33(7H, m), 2.47˜3.05(2H, m), 3.32˜3.51(1H, m),3.63˜3.85(1H, m), 3.85˜4.12(3H, m), 4.16˜4.30(1H, m), 4.43˜4.51(1H, m),5.15˜5.29(4H, m), 7.45˜7.56(4H, m), 8.21(4H, d, J=9 Hz)

Production Example 22

(2S,4S)-4-Acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-3-yl-hydroxymethyl]-pyrrolidinediastereomer B ##STR58##

The reaction of Production Example 19 was repeated by using(2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxy-carbonyl)piperidine-3-ylhydroxymethyl]pyrrolidinediastereomer B (757 mg, 1.36 mmol) to thereby give the target compound(555 mg, 66.2%).

NMR(CDCl₃) δ: 1.20˜1.56(3H, m), 1.67˜1.80(1H, m), 1.90˜2.47(3H, m),2.35(3H, s), 2.64˜2.98(2H, m), 3.02˜3.18(1H, m), 3.66˜4.30(6H, m),5.13˜5.32(4H, m), 7.45˜7.59(4H, m), 8.18˜8.28(4H, m)

Example 12

p-Nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxy-carbonyl)piperidine-3-ylhydroxymethyl]-pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylatediastereomer B ##STR59##

The reaction of Example 10 was repeated by using(2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-3-ylhydroxymethyl]-pyrrolidinediastereomer B (540 mg, 0.877 mmol) to thereby give the target compound(589 mg, 73.2%).

NMR(CDCl₃) δ: 1.15˜2.48(7H, m), 1.28(3H, d, J=7 Hz), 1.38(3H, d, J=6Hz), 2.50˜2.99(2H, m), 3.11˜3.62(3H, m), 3.28(1H, dd, J=2, 7 Hz),3.74˜4.32(6H, m), 4.25(1H, dd, J=2, 9 Hz), 5.13˜5.32(5H, m), 5.50(1H, d,J=13 Hz), 7.42˜7.56(4H, m), 7.65(2H, d, J=9 Hz), 8.15˜8.27(6H, m)

Example 13

(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-1-methyl-2-[(2S,4S)-2-piperidine-3-ylhydroxymethyl)pyrrolidin-4-ylthio]-carbapen-2-em-3-carboxylicacid acetate diastereomer B ##STR60##

The reaction of Example 11 was repeated by using p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyl-oxycarbonyl)-piperidine-3-ylhydroxymethyl)pyrrolidin-4-ylthio]-carbapen-2-em-3-carboxylatediastereomer B (560 mg, 0.610 mmol) to thereby give the target compound(119 mg. 40.0%).

NMR(D₂ O) δ: 1.17(3H, d, J=7 Hz), 1.23(3H, d, J=6 Hz), 1.28˜1.41(1H, m),1.58˜1.73(1H, m), 1.76˜1.93(3H, m), 1.86(3H, s), 1.93˜2.03(2H, m),2.60(1H, td, J=8, 14 Hz), 2.83(1H, t, J=12 Hz), 2.86(1H, dt, J=3, 13Hz), 3.26˜3.40(4H, m), 3.42(1H, dd, J=2, 6 Hz), 3.61(1H, dd, J=7, 12Hz), 3.77˜3.85(2H, m), 3.92˜4.02(1H, m), 4.15˜4.24(2H, m)

M.S. (m/e): 426(MH⁺)

Production Example 23

(2S,4R)-2-(N-Benzylpiperidin-4-ylhydroxymethyl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-pyrrolidine##STR61##

At 55° C., a catalytic amount of iodine and ethyl bromide (0.075 ml)were added to a suspension (4 ml) of a magnesium powder (486 mg, 20.0mmol) in tetrahydrofuran. Into the obtained solution was dropped, underheating, a solution of N-benzyl-4-chloropiperidine (4.61 g, 20.0 mmol)in tetrahydrofuran (25 ml) over 20 minutes. After the completion of thedropping, the resulting mixture was heated under reflux for 1 hour. Thesolution was cooled by allowing it to stand and then dropped into asolution of(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethyl-siloxy-2-formylpyrrolidine(6.58 g, 20.0 mmol) in tetrahydrofuran (60 ml) cooled to -30° C. Afterthe completion of the dropping, the reaction mixture was stirred for 1hour while slowly returning to room temperature. After adding asaturated aqueous solution of ammonium chloride (20 ml) to the reactionmixture, the tetrahydrofuran was distilled off under a reduced pressure.Then, ethyl acetate (120 ml) was added to the residue. The organic layerwas washed successively with water, 1 N hydrochloric acid, water, a 10%aqueous solution of potassium carbonate, water and a saturated aqueoussolution of sodium chloride. The organic layer was dried over anhydrousmagnesium sulfate and concentrated under a reduced pressure. The residuewas subjected to silica gel chromatography [Wakogel C-200, 100 g,hexane-ethyl acetate (3:1 to 1:1)] to thereby give the target compound(3.40 g, 33.7%).

NMR(CDCl₃) δ: 0.05(6H, s), 0.86(9H, s), 1.20˜1.33(1H, m), 1.36˜2.00(8H,m), 1.46(9H, s), 2.84˜2.99(2H, m), 3.20˜3.31(1H, m), 3.21(1H, dd, J=4,12 Hz), 3.48(1H, d, J=16 Hz), 3.49(1H, d, J=16 Hz), 3.56(1H, d, J=12Hz), 4.10(1H, q, J=8 Hz), 4.25(1H, brs), 5.02(1H, brs), 7.19˜7.36(5H, m)

Production Example 24

(2S,4R)-N-tert-Butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[N-(p-nitrobenzyloxycarbonyl)-piperidine-4-ylhydroxy-methyl]-pyrrolidine##STR62##

To a solution of(2S,4R)-2-(N-benzylpiperidin-4-ylhydroxymethyl]-N-tert-butoxycarbonyl-4-tert-butyldimethyl-siloxypyrrolidine(748 mg, 1.48 mmol) in methanol (10 ml) were added ammonium formate(0.47 g, 7.5 mmol) and 10% palladium-carbon (0.75 g) and the obtainedmixture was heated under reflux for 15 minutes. After filtering off thecatalyst through celite, the solvent was distilled off. Then a 5%aqueous solution of potassium carbonate (15 ml) was added to the residuefollowed by extraction with ethyl acetate. The organic layer was washedwith a saturated aqueous solution of sodium chloride, dried overanhydrous magnesium sulfate and concentrated under a reduced pressure.To the solution of the residue thus obtained in tetrahydrofuran (10 ml)were added, under ice-cooling, triethylamine (0.18 ml, 1.3 mmol) andp-nitrobenzyl chloroformate (255 mg, 1.18 mmol) and the mixture wasstirred at the same temperature for 6 hours. After adding ethyl acetate(60 ml) to the reaction mixture, the organic layer was washedsuccessively with water, 1 N hydrochloric acid, water, a saturatedaqueous solution of sodium hydrogen-carbonate and a saturated aqueoussolution of sodium chloride. The organic layer was dried over anhydrousmagnesium sulfate and concentrated under a reduced pressure to therebygive the target compound as a crude product (624 mg, 71.1%).

NMR(CDCl₃) δ: 0.05(6H, s), 0.86(9H, s), 1.40˜1.85(6H, m), 1.46(9H, s),1.90˜2.02(1H, m), 2.60˜2.92(2H, m), 3.22(1H, dd, J=4, 12 Hz), 3.31(1H,d, J=8 Hz), 3.61(1H, d, J=12 Hz), 4.12(1H, q, J=8 Hz), 4.16˜4.37(3H, m),5.15˜5.40(3H, m), 7.50(2H, d, J=8 Hz), 8.21(2H, d, J=8 Hz)

Production Example 25

(2S,4R)-4-Hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-4-ylhydroxymethyl)-pyrrolidine##STR63##

Under ice-cooling, trifluoroacetic acid (20 ml) was added to a solutionof crude(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-4-ylhydroxymethyl]-pyrrolidine(3.89 g. 6.56 mmol) in methylene chloride (20 ml) and the obtainedmixture was stirred at the same temperature for 40 minutes and then atroom temperature for additional 40 minutes. Next, the solvent andtrifluoro-acetic acid were distilled off under a reduced pressure. A 10%aqueous solution of potassium carbonate (30 ml) was added to the residuefollowed by extraction with ethyl acetate. The organic layer was washedwith a saturated aqueous solution of sodium chloride, dried overanhydrous magnesium sulfate and concentrated. The residue thus obtainedwas dissolved in tetrahydrofuran (30 ml). To the solution thus obtainedwas added, under ice-cooling, a solution of triethylamine (0.94 ml, 6.7mmol) and p-nitrobenzyl chloroformate (1.39 g, 6.45 mmol) intetrahydrofuran (15 ml) and the resulting mixture was stirred at thesame temperature for 10 minutes and then at room temperature for anadditional 2.5 hours. After adding ethyl acetate (120 ml) to thereaction mixture, the organic layer was washed successively with water,1 N hydrochloric acid, water, a saturated aqueous solution of sodiumhydrogen-carbonate and a saturated aqueous solution of sodium chloride.The organic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was dissolved intetrahydrofuran (40 ml). To the obtained solution was added, underice-cooling, a 1 M solution of tetrabutyl-ammonium fluoride intetrahydrofuran (8.9 ml, 8.9 mmol) and the mixture was stirred at thesame temperature for 15 minutes and then at room temperature for anadditional 2 hours. After distilling off the solvent under a reducedpressure, ethyl acetate (80 ml) was added to the residue. The organiclayer was washed successively with water, 1 N hydrochloric acid, a50%-saturated aqueous solution of sodium chloride, a saturated aqueoussolution of sodium hydrogen-carbonate and a saturated aqueous solutionof sodium chloride, dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography (Wakogel C-200, 35 g, ethyl acetate) tothereby give the target compound (2.00 g, 54.6%).

NMR(CDCl₃) δ: 1.45˜1.76(5H, m), 1.77˜1.95(1H, m), 2.06˜2.16(1H, m),2.60˜2.90(2H, m), 3.30˜3.50(2H, m), 3.82(1H, d, J=12 Hz), 4.15˜4.35(3H,m), 4.46(1H, brs), 4.65˜4.86(1H, m), 5.15˜5.28(4H, m), 7.50(2H, d, J=8Hz), 7.51(2H, d, J=8 Hz), 8.22(4H, d, J=8 Hz)

Production Example 26

(2S,4S)-4-Acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-4-yl-hydroxymethyl]-pyrrolidine##STR64##

Under ice-cooling, triethylamine (0.68 ml, 4.9 mmol) and methanesulfonylchloride (0.36 ml, 4.7 mmol) were added to a solution of(2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-4-ylhydroxymethyl]-pyrrolidine(1.99 g, 3.57 mmol) in methylene chloride (20 ml) and the obtainedmixture was stirred at the same temperature for 2 hours. Then, thereaction mixture was washed successively with water, a saturated aqueoussolution of sodium hydrogencarbonate and a saturated aqueous solution ofsodium chloride. The organic layer was dried over anhydrous magnesiumsulfate and concentrated under a reduced pressure. To the solution ofthe obtained residue in dimethylformamide (20 ml) was added potassiumthioacetate (1.22 g, 10.7 mmol) and the mixture was stirred at 70° C.for 3 hours. After distilling off the solvent under a reduced pressure,ethyl acetate (50 ml) was added to the residue. The organic layer waswashed successively with water, a saturated aqueous solution of sodiumhydrogen-carbonate, water and a saturated aqueous solution of sodiumchloride. Next, the organic layer was dried over anhydrous magnesiumsulfate and concentrated under a reduced pressure. The residue wassubjected to silica gel chromatography [Wakogel C-200, 20 g, ethylacetate-hexane (1:1 to 2:1)] to thereby give the target compound (1.57g, 71.4%).

NMR(CDCl₃) δ: 1.48˜2.10(6H, m), 2.36(3H, s), 2.45˜2.60(1H, m),2.63˜2.93(2H, m), 3.18(1H, dd, J=9, 11 Hz), 3.42˜3.51(1H, m),3.77˜3.88(1H, m), 4.10(1H, q, J=8 Hz), 4.15˜4.40(3H, m), 4.50˜4.75(1H,m), 5.15˜5.29(4H, m), 7.46˜7.55(4H, m), 8.17˜8.26(4H, m)

Example 14

p-Nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitro-benzyloxy-carbonyl)piperidine-4-ylhydroxymethyl]-pyrrolidin-4-ylthio}-carbapen-2-em-3-carboxylate##STR65##

Under ice-cooling, a 1 N aqueous solution of sodium hydroxide (2.66 ml,2.66 mmol) was added to a solution of(2S,4S)-4-acetylthio-N-(p-nitrobenzyloxy-carbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidine-3-ylhydroxymethyl]-pyrrolidine(1.56 g, 2.53 mmol) in a mixture of methanol (15 ml) withtetrahydrofuran (15 ml) and the obtained mixture was stirred at the sametemperature for 30 minutes. After adding 1 N hydrochloric acid (2.75 ml,2.75 mmol) to the reaction mixture, the solvent was distilled off undera reduced pressure and ethyl acetate (60 ml) was added to the residue.Then, the organic layer was washed successively with water and asaturated aqueous solution of sodium chloride, dried over anhydrousmagnesium sulfate and concentrated under a reduced pressure. To thesolution of the obtained residue in acetonitrile (20 ml) were added,under ice-cooling, p-nitrobenzyl(1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methylcarbapen-2-em-3-carboxylate(1.46 g, 2.46 mmol) and diisopropyl-ethylamine (0.45 ml, 2.6 mmol). Theobtained mixture was stirred at the same temperature for 40 minutes andthen at room temperature for an additional 40 minutes. After addingethyl acetate (100 ml) to the reaction mixture, the organic layer waswashed successively with water, a 50%-saturated aqueous solution ofsodium chloride and a saturated aqueous solution of sodium chloride.Next, the organic layer was dried over anhydrous magnesium sulfate andconcentrated under a reduced pressure. The residue was subjected tosilica gel chromatography (Wakogel C-200, 40 g, 1% methanol-chloroform)to thereby give the target compound (1.03 g, 44.6%).

NMR(CDCl₃) δ: 1.28(3H, d, J=8 Hz), 138(3H, d, J=7 Hz), 1.45˜2.00(6H, m),2.47˜260(1H, m), 2.63˜2.93(2H, m), 3.18˜3.40(2H, m), 3.28(1H, dd, J=2, 7Hz), 3.40˜3.63(2H, m), 4.05˜4.40(6H, m), 5.15˜5.32(5H, m), 5.50(1H, d,J=13 Hz), 7.47˜7.55(4H, m), 7.65(2H, d, J=8 Hz), 8.18˜8.27(6H, m)

Example 15

(1R,5S,6S)-6-[(R)-1-Hydroxyethyl]-1-methyl-2-[(2S,4S)-2-piperidine-4-ylhydroxymethyl)pyrrolidin-4-ylthio]carbapen-2-em-3-carboxylicacid acetate ##STR66##

To a solution of p-nitrobenzyl(1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-{(2S,4S)-N-(p-nitro-benzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)-piperidine-4-ylhydroxymethyl]pyrrolidin-4-ylthio}carbapen-2-em-3-carboxylate(1.03 g, 1.12 mmol) in a mixture of a 0.1 M phosphate buffer (pH 7.0, 20ml) with tetrahydrofuran (20 ml) was added 20% palladiumhydroxide/carbon (1.03 g) and hydrogenation was effected by using aPerlman shaker under a moderate pressure (4 kg/cm²) at room temperaturefor 1.5 hours. After filtering off the catalyst through celite, theorganic solvent was distilled off from the filtrate under a reducedpressure. The remaining aqueous layer was washed with diethyl ether andthen the organic solvent was distilled off again from the aqueous layerunder a reduced pressure. The pH value of the remaining aqueous layerwas adjusted to 7.5 by using a 1 N aqueous solution of sodium hydroxide.After filtering off the insoluble matters, the filtrate was subjected toreversed phase silica gel chromatography (YMC SH-343-7 AM ODS, 0.005 Macetic acid-2% aqueous solution of methanol). The target fraction wasconcentrated and then freeze-dried to thereby give the target compound(178 mg, 32.7%).

NMR(D₂ O) δ: 1.17(3H, d, J=7 Hz), 1.24(3H, d, J=6 Hz), 1.50˜1.90(6H, m),1.86(3H, s), 2.65(1H, td, J=8, 14 Hz), 2.97(2H, q, J=13 Hz),3.27˜3.38(2H, m), 3.38˜3.50(3H, m), 3.61(1H, dd, J=6, 12 Hz),3.72˜3.83(2H, m), 3.95˜4.05(1H, m), 4.14˜4.24(2H, m)

M.S. (m/e): 426(MH⁺)

We claim:
 1. A carbapenem compound represented by the following formula(I), or a salt thereof: ##STR67## wherein the ring A represents a 3- to7-membered saturated or partially unsaturated heterocycle containingonly nitrogen as heteroatoms and optionally having a ring substituentother than R⁶ and selected from the group consisting of a hydrogen atom,hydroxyl, mercapto, halogen, cyano, azido, alkyl, alkenyl, alkynyl,alkoxy, halogenoalkyl, guanidino, formimidoyl, acetimidoyl, carbamoyl,thiocarbamoyl, carbamoylalkyl, carbamido, alkanoyl, amino, alkylamino,dialkylamino, aminoalkyl, carboxy, alkoxycarbonyl, alkoxycarbonylalkyl,aminoalkylaminoalkyl, alkylcarbonyloxy, cycloalkyl, cycloalkenyl,phenyl, alkylthio, phenylthio, benzyl, benzoyl and halogenoaryl; R¹represents hydrogen or methyl; R² and R⁵ are the same or different andeach represents hydrogen or a protecting group of the hydroxyl groupselected from the group consisting of trimethylsilyl, t-butyldimethylsilyl, methoxymethyl, 2-methoxyethoxymethyl, tetrahydropyranyl,benzyl, p-methoxybenzyl, 2,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, trityl, formyl, acetyl, t-butoxy-carbonyl,2-iodoethoxycarbonyl, 2,2,2-trichloroethoxy-carbonyl,2-propenyloxycarbonyl, 2-chloro-2-propenyloxycarbonyl,3-methoxycarbonyl-2-propenyloxycarbonyl, 2-methyl-2-propenyloxycarbonyl,2-butenyloxycarbonyl, cinnamyloxycarbonyl, benzyloxycarbonyl,p-methoxybenzyloxycarbonyl, o-nitrobenzyl-oxycarbonyl andp-nitrobenzyloxycarbonyl; R³ represents hydrogen or a protecting groupof the carboxyl group selected from the group consisting of methyl,ethyl, isopropyl, t-butyl, 2-iodoethyl, 2,2,2-trichloroethyl,methoxymethyl, ethoxymethyl, isobutoxymethyl, butyrloxymethyl,pivaloyloxy-methyl, 1-methoxycarbonyloxyethyl, 1-ethoxycarbonyloxyethyl,benzyl, p-methoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, benzhydryl andphthalidyl; R⁴ represents hydrogen, lower alkyl or a protecting group ofthe amino group selected from the group consisting of formyl, acetyl,chloroacetyl, dichloroacetyl, propionyl, phenylacetyl, phenoxyacetyl,thienylacetyl, benzyloxycarbonyl, t-butoxy carbonyl,p-nitrobenzyloxycarbonyl, 2,2,2-trichloroethyl, trityl, p-methoxybenzyl,p-nitrobenzyl, diphenylmethyl, pivaloyloxymethyl, trimethylsilyl,t-butyl dimethylsilyl, benzylidene, salicylidene, p-nitrobenzylidene,m-chlorobenzylidene, 3,5-di(t-butyl)-4-hydroxybenzylidene and3,5-di(t-butyl)benzylidene; R⁶ represents: (1) hydrogen, (2) loweralkyl, optionally substituted by an optionally protected hydroxy, acarbamoyl, formimidoyl, acetimimidoyl or ##STR68## wherein R⁷ and R⁸ arethe same or different and each represents hydrogen, lower alkyl, or aprotecting group of the amino group, or (3) a protecting group of theamino group or a protecting group of the imino group; and m is 0 or 1,wherein the hydroxy is protected by the hydroxyl protecting groupdescribed above and the amino and imino group are protected by the aminoprotecting group described above.
 2. A carbapenem compound representedby the following formula (II), or a salt thereof: ##STR69## wherein thering A represents a 3- to 7-membered saturated or partially unsaturatedheterocycle containing only nitrogen as heteroatoms and optionallyhaving a ring substituent other than R⁶ and selected from the groupconsisting of a hydrogen atom, hydroxyl, mercapto, halogen, cyano,azido, alkyl, alkenyl, alkynyl, alkoxy, halogenoalkyl, guanidino,formimidoyl, acetimidoyl, carbamoyl, thiocarbamoyl, carbamoylalkyl,carbamido, alkanoyl, amino, alkylamino, dialkylamino, aminoalkyl,carboxy, alkoxycarbonyl, alkoxycarbonylalkyl, aminoalkylaminoalkyl,alkylcarbonyloxy, cycloalkyl, cycloalkenyl, phenyl, alkylthio,phenylthio, benzyl, benzoyl and halogenoaryl; R¹ represents hydrogen ormethyl; R² and R⁵ are the same or different and each represents hydrogenor a protecting group of the hydroxyl group selected from the groupconsisting of trimethylsilyl, t-butyl dimethylsilyl, methoxymethyl,2-methoxyethoxymethyl, tetrahydropyranyl, benzyl, p-methoxybenzyl,2,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, trityl, formyl,acetyl, t-butoxycarbonyl, 2-iodoethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-propenyloxycarbonyl,2-chloro-2-propenyloxycarbonyl, 3-methoxycarbonyl-2-propenyloxycarbonyl,2-methyl-2-propenyloxycarbonyl, 2-butenyloxycarbonyl,cinnamyloxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl,o-nitrobenzyloxycarbonyl and p-nitrobenzyloxycarbonyl; R³ representshydrogen or a protecting group of the carboxyl group selected from thegroup consisting of methyl, ethyl, isopropyl, t-butyl, 2-iodoethyl,2,2,2-trichloroethyl, methoxymethyl, ethoxymethyl, isobutoxymethyl,butyrloxymethyl, pivaloyloxy-methyl, 1-methoxycarbonyloxyethyl,1-ethoxycarbonyloxyethyl, benzyl, p-methoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, benzhydryl and phthalidyl; R⁴ represents hydrogen, loweralkyl or a protecting group of the amino group selected from the groupconsisting of formyl, acetyl, chloroacetyl, dichloroacetyl, propionyl,phenylacetyl, phenoxyacetyl, thienylacetyl, benzyloxycarbonyl, t-butoxycarbonyl, p-nitrobenzyloxycarbonyl, 2,2,2-trichloroethyl, trityl,p-methoxybenzyl, p-nitrobenzyl, diphenylmethyl, pivaloyloxy-methyl,trimethylsilyl, t-butyl dimethylsilyl, benzylidene, salicylidene,p-nitrobenzylidene, m-chlorobenzylidene,3,5-di(t-butyl)-4-hydroxybenzylidene and 3,5-di(t-butyl)benzylidene; R⁶represents: (1) hydrogen, (2) lower alkyl, optionally substituted by anoptionally protected hydroxy, a carbamoyl, formimidoyl, acetimimidoyl or##STR70## wherein R⁷ and R⁸ are the same or different and eachrepresents hydrogen, lower alkyl, or a protecting group of the aminogroup, or (3) a protecting group of the amino group or a protectinggroup of the imino group; and m is 0 or 1, wherein the hydroxy isprotected by the hydroxyl protecting group described above and the aminoand imino group are protected by the amino protecting group describedabove.
 3. A compound as claimed in claim 1, or a salt thereof, whereinR¹ is methyl, R², R³ , R⁴, R⁵ and R⁶ are each hydrogen and m is
 0. 4. Acompound as claimed in claim 1, or a salt thereof, wherein R¹ is methyl,R², R³, R⁴, R⁵ and R⁶ are each hydrogen and m is
 1. 5. The compoundrepresented by the following formula, or a salt thereof: ##STR71## 6.The compound represented by the following formula, or a salt thereof: 7.The compound represented by the following formula, or a salt thereof: 8.The compound represented by the following formula, or a salt thereof: 9.A process for producing a compound as claimed in claim 1, or a saltthereof, which comprises reacting a compound represented by thefollowing formula (III) or a reactive derivative thereof: wherein R¹represents hydrogen or methyl; R² represents hydrogen or the protectinggroup of the hydroxyl group; and R³ represents hydrogen or theprotecting group of the carboxyl group, with a mercaptan represented bythe following general formula (IV): ##STR72## wherein the ring Arepresents a 3- to 7-membered saturated or partially unsaturatedheterocycle containing only nitrogen as heteroatoms and optionallyhaving a ring substituent other than R⁶ and selected from the groupconsisting of a hydrogen atom, hydroxyl, mercapto, halogen, cyano,azido, alkyl, alkenyl, alkynyl, alkoxy, halogenoalkyl, guanidino,formimidoyl, acetimidoyl, carbamoyl, thiocarbamoyl, carbamoylalkyl,carbamido, alkanoyl, amino, alkylamino, dialkylamino, aminoalkyl,carboxy, alkoxycarbonyl, alkoxycarbonylalkyl, aminoalkylaminoalkyl,alkylcarbonyloxy, cycloalkyl, cycloalkenyl, phenyl, alkylthio,phenylthio, benzyl, benzoyl and halogenoaryl; R⁴ represents hydrogen,lower alkyl or the protecting group of the amino group; R⁵ representshydrogen or the protecting group of the hydroxyl group; R⁶ represents:(1) hydrogen, (2) lower alkyl, optionally substituted by the optionallyprotected hydroxy, the carbamoyl, formimidoyl, acetimidoyl or ##STR73##wherein R⁷ and R⁸ are the same or different and each representshydrogen, lower alkyl, or the protecting group of the amino group, or(3) the protecting group of the amino group or the protecting group ofthe imino group; and m is 0 or 1, optionally followed by the deblockingreaction.
 10. A compound as claimed in claim 1, wherein the ring A isselected from the group consisting of aziridine, azetidine, pyrrolidine,piperidine, homopiperidine, imidazolidine, pyrazolidine, piperazine,pyrroline, imidazoline and pyrazoline.
 11. In a method for preventing ortreating an illness caused by a bacteria, the improvement comprisingadministering a pharmacologically efficacious dose of a compound asclaimed in claim 1, or a salt thereof, to a patient.
 12. A compound asclaimed in claim 2 or a salt thereof, wherein R¹ is methyl, R², R³, R⁴,R⁵ and R⁶ are each hydrogen and m is
 1. 13. A medicinal compositionwhich comprises a pharmacologically efficacious amount of a compound asclaimed in claim 1, or a salt thereof, and pharmacologically acceptablecarrier(s).
 14. A compound as claimed in claim 2 or a salt thereof,wherein R¹ is methyl, R², R³, R⁴, R⁵ and R⁶ are each hydrogen and m is0.